CC-1065 analogs having two CPI subunits

ABSTRACT

PCT No. PCT/US89/03329 Sec. 371 Date Mar. 8, 1991 Sec. 102(e) Date Mar. 8, 1991 PCT Filed Aug. 7, 1989 PCT Pub. No. WO90/02746 PCT Pub. Date Mar. 22, 1990This invention concerns chemical compounds of general Formula ICPI1-R5-T-R&#39;5-CPI2I The compounds of Formula I are useful as uv light absorbers, antibacterial agents, and are particularly useful as antitumor agents. Representative compounds of Formula I have been shown to possess useful ranges of antitumor activity in standard laboratory animal tests.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is the national phase of PCT/US89/03329, filed 7 Aug.1989; which is a continuation of Ser. No. 07/243,350, filed 12 Sep.1988, now abandoned.

BACKGROUND OF THE INVENTION

Antibiotic CC-1065,(7bR,8aS)-7-[[1,6-dihydro-4-hydroxy-5-methoxy-7-[(4,5,8,8a-tetrahydro-7-methyl-4-oxocyclopropa[c]pyrrolo[3,2-e]indol-2(1H)-yl)carbonyl]benzo[1,2-b:4,3-b']dipyrrol-3(2H)-yl]carbonyl]1,6-dihydro-4-hydroxy-5-methoxy-benzo[1,2-b:4,3-b']dipyrrole-3(2H)-carboxamide,is disclosed and claimed in L. J. Hanka et al. U.S. Pat. No. 4,169,888together with a process for preparing antibiotic CC-1065 by aerobicfermentation procedures, and recovering antibiotic CC-1065 therefrom.

In The Journal of Antibiotics, 1985, 38, 746, D. G. Martin et alreported that acetic acid adds across thespirocyclopropylcyclohexadienyl (SCPCH) system of CC-1065 to produce thephenolic, acetic acid product (AAP),7-[[7-[[1-[(acetyloxy)methyl]-1,6-dihydro-5-hydroxy-8-methylbenzo[1,2-b:4,3-b']dipyrrol-3(2H)-yl]carbonyl]-1,6-dihydro-4-hydroxy-5-methoxybenzo[1,2-b:4,3-b']-dipyrrol-3(2H)-yl]carbonyl]-1,6-dihydro-4-hydroxy-5-methoxy-benzo[1,2-b:4,3-b']dipyrrole-3(2H)-carboxamide.AAP was tested in vitro and in vivo and found to be less potent thanCC-1065 by a factor of 10³ to 10⁴ depending upon the particular testsystem and therefore tended to divert attention from adducts of theSCPCH system as useful antitumor agents or as prodrugs to CC-1065analogs.

In J. Am. Chem. Soc., 103, No. 18, 1981, W. Wierenga published a"Synthesis of the Left-Hand Segment of the Antitumor Agent CC-1065".

EP Application 0 154 445 (published Nov. 9, 1985) discloses variousanalogs of antibiotic CC-1065, including compounds of formula EP-I andEP-II (see General Formula chart of EP 0154 445), wherein R₁ in formulaEP-II is CH₃ --, --CH₂ Ph, CH═CHCH₂ --, --CH₂ SCH₃, --CH₂ OCH₃, --CH₂OCH₂ CH₂ OCH₃, --CH₂ CCl₃, --CH₂ CH₂ Si(R₂)₃, or H, where Ph is phenyl;R is alkyl(C₁ -C₅), phenyl, or H; R₂ ' is C₁ to C₅ -alkyl, phenyl orhydrogen, and is not necessarily the same as R in one compound; R₃ isalkyl(C₁ -C₅), phenyl, or H; and X is Cl, Br, I or OSO₂ R₄₀, where R₄₀is C₁ to C₅ -alkyl, phenyl, tolyl, bromophenyl, nitrophenyl, ortrifluromethyl. The O-protected compounds of formula EP-II arechemically stable and only removable under specific chemical conditions.However, when the compounds of formula EP-II are O-deprotected, they canbe cyclized to yield the compounds of EP-I.

EP Application 0 154 445 also discloses CPI dimers joined by--CO--(CH₂)_(n1) --CO-- where n₁ is 2-12 and CPI dimers joined by thetether --C(O)--(--R₁₁ --)--C(O)--X₇ --(CH₂ CH₂ --X₇)n4--C(O)--(--R₁₁--)--C(O)-- where R₁₁ =CH₂ CH₂, CH═CH; and X₇ --O, NH, and n4=1-4, andthe HCl and MeI salts for X₇ =NH.

Additional dimers of CPI prodrugs joined by --CO--(CH₂)_(n1) --CO--where n₁ is 2-2 and CPI dimers Joined by the tether --C(O)--(--R₁₁--)--C(O)--X₇ --(--CH₂ CH₂ --X₇)n4--C(O)--(--R₁₁)--C(O)-- where R₁₁ =CH₂CH₂, CH═CH; and X₇ =O, NH, and n4=1-4, and the HCl and MeI salts for X₇=NH are disclosed in U.S. patent application Ser. No. 944,633, filed 19Dec. 1986, now abandoned, and PCT/U.S. application Ser. No. 87/03227,filed 11 Dec. 1987, published 14 Jul. 1988.

Various oral and poster presentations of material in U.S. patentapplication Ser. No. 944,633, filed 19 Dec. 1986, have been made.

SUMMARY OF THE INVENTION

This invention provides some new synthetically obtained compounds offormula I (see General Formulae Chart), as defined hereinafter, whichare useful as uv light absorber substances, or as chemicalintermediates. Representative formula I compounds have also been shownto possess useful ranges of antitumor activity in standard laboratoryanimal tests. The compounds of this invention are obtained by chemicalprocesses shown in Schemes 1-6 and detailed in the examples.

DETAILED DESCRIPTION OF THE INVENTION

More specifically, this invention provides new chemical compounds ofgeneral Formula I (see GENERAL FORMULA sheet)

wherein CPI₁ and CPI₂, being the same or different, are selected fromFormula A or B (see GENERAL FORMULA sheet);

wherein W is selected from C₁ -C₅ alkyl, phenyl or hydrogen;

wherein X is selected from azido, a halogen atom, cyanate, thiocyanate,isocyanate, thioisocyanate, phosphate diester (--O--PO(OR)₂), phosphonyl(--O--PO₂ R), thiophosphonyl (--O--PSOR), sulfinyl (--O--SOR) orsulfonyl (--O--SO₂ R);

wherein Y is selected from hydrogen, --C(O)R, --C(S)R, --C(O)OR₁,--S(O)₂ R₁, --C(O)NR₂ R₃, --C(S)NR₂ R₃, or --C(O)NHSO₂ R₄ ;

wherein Z is selected from the group consisting of C₁ -C₅ alkyl, phenylor hydrogen:

wherein R is selected from the group consisting of C₁ -C₂₀ alkyl; C₂ -C₆alkenyl; C₂ -C₆ alkynyl; phenyl optionally substituted with one, 2 or 3C₁ -C₄ alkyl, C₁ -C₃ alkoxy, halo, C₁ -C₃ alkylthio, trifluoromethyl, C₂-C₆ dialkylamino, or nitro; naphthyl optionally substituted with one or2 C₁ -C₄ alkyl, C₁ -C₃ alkoxy, halo, trifluromethyl, C₂ -C₆dialkylamino, C₁ -C₃ alkylthio or nitro;

wherein R₁ is selected from C₁ -C₂₀ alkyl or phenyl optionallysubstituted with one, 2 or 3 C₁ -C₄ alkyl, C₁ -C₃ alkoxy, halo, C₁ -C₃alkylthio, trifluoromethyl, C₂ -C₆ dialkylamino, or nitro;

wherein R₂ and R₃, being the same or different, are selected fromhydrogen, C₁ -C₂₀ alkyl, or phenyl optionally substituted with one, 2 or3 C₁ -C₄ alkyl, C₁ -C₃ alkoxy, halo, C₁ -C₃ alkylthio, trifluoromethyl,C₂ -C₆ dialkylamino, or nitro; with the proviso that both R₂ and R₃ cannot be phenyl or substituted phenyl;

wherein R₄ is selected from C₁ -C₁₀ alkyl; phenyl optionally substitutedwith one, 2 or 3 C₁ -C₄ alkyl, C₁ -C₃ alkoxy, halo, C₁ -C₃ alkylthio,trifluoromethyl, C₂ -C₆ dialkylamino, or nitro; naphthyl optionallysubstituted with one or 2 C₁ -C₄ alkyl, C₁ -C₃ alkoxy, halo,trifluromethyl, C₂ -C₆ dialkylamino, C₁ -C₃ alkylthio or nitro;

wherein T is a tether linkage selected from the group consisting of:

a) aminocarbonyl (--NHC(O)--);

b) carbonylamino (--C(O)NH--);

c) carbonyloxy (--C(O)O--);

d) oxycarbonyl (--OC(O)--);

e) amino-tether-amino of the formula --NR₁₃ --T'--NR₁₄ -- where R₁₃ andR₁₄, being the same or different, are hydrogen, or C₁ -C₈ alkyl, or whentaken together are --(CH₂)_(n) -- where n is! 2 or 3; where T' isselected from the group consisting of carbonyl (--C(O)--), dicarbonyl(--C(O)C(O)--), (--C(O)(CH₂)_(n) C(O)--), where n is 1 to 5,(--C(O)PhC(O)--), where Ph is 1,3- or 1,4-phenylene, or a group of theformula --C(O)-het-C(O)--, where -het- is as defined below; or

f) --C(O)-het-C(O)--, when R₅ and R'₅ are both a direct bond, wherein-het- is a fused mono-, di-, or tricyclic heteroaryl of 5 to 12 members,containing one, two, or three heteroatoms selected from the groupconsisting of oxygen, nitrogen or sulfur, optionally substituted withone or 2 C₁ -C₄ alkyl, C₁ -C₃ alkoxy, halo, C₁ -C₃ alkylthio,trifluoromethyl, C₂ -C₆ dialkylamino, or nitro;

wherein R₅ and R'₅, being the same or different, are selected from adirect bond or a carbonyl acyl group selected from the group consistingof a compound of formula (ii), (vi), (viii), (x), (xi), (xvii), (xviia),(xviib), (xviii), (xix), (xx), (xxi), (xxii), (xxiii), (xxiv), (xxv),(xxvi) as defined in Chart C where the indicated free valence bonded tothe carbonyl carbon atom is bonded to the indicated free valence of thenitrogen (N--) in CPI₁ or CPI₂ and the other indicated free valencerepresents a bond to the tether group T.

CPI₁ and CPI₂, being the same or different, are preferably1-(chloromethyl)-1,6-dihydro-8-methyl-5-hydroxy-benzo[1,2-b:4,3-b']dipyrrole-3(2H)-ylor4,5,8,8a-tetrahydro-7-methyl-4-oxocyclopropa[c]-pyrrolo(3,2-e)indol-2(1H)-yl.

W is preferably methyl.

X is preferably halogen, more preferably chloro or bromo.

Y is preferably --COR, wherein R is selected from C₁ -C₁₀ alkyl; phenyloptionally substituted with one, 2 or 3 C₁ -C₄ alkyl, C₁ -C₃ alkoxy,halo, C₁ -C₃ alkylthio, trifluoromethyl, C₂ -C₆ dialkylamino, or nitro;--C(O)NHSO₂ R₄ ; or --C(O)NR₂ R₃.

Z is preferably hydrogen.

T is preferably an amide (aminocarbonyl); carbonylamino (--C(O)NH--); oran amino-tether-amino of the formula --NR₁₃ --T'--NR₁₄ -- where R₁₃ andR₁₄ are hydrogen and where T' is selected from carbonyl (--C(O)--) or agroup of the formula --C(O)-het-C(O)--, where -het- is a heteroarylselected from Supplemental Formula Chart, especially preferred arepyrrol-2,5-diyl, fur-2,5-diyl, indol-2,5-diyl, benzofuran-2,5-diyl or3,6-dihydrobenzo[1,2-b:4,3-b']dipyrrol-2,7-diyl. The indicated freevalences of -het- are bonded to the carbonyl carbon atoms(--C(O)-het-C(O)--).

R₅ is preferably 2-carbonylindole-5-yl,2-carbonyl-6-hydroxy-7-methoxyindol-5-yl,2-carbonyl-1,2,3,6-tetrahydrobenzo[1,2-b:4,3b']dipyrrol-6-yl,2-carbonyl-3-hydroxy-4-methoxy-1,2,3,6-tetrahydrobenzo[1,2-b:4,3-b']dipyrrol-6-yl.

Halogen atom (halo) refers to a bromo, chloro, iodo or fluoro atom.

Examples of C₁ -C₂₀ alkyl are methyl, ethyl, propyl, butyl and the like,including isomeric forms thereof. Examples of C₁ -C₃ alkoxy are methoxy,ethoxy, propoxy and isomeric forms thereof. Examples of C₂ -C₆dialkylamino are dimethylamino, diethylamino, methylethylamino,dipropylamino and ethylpropylamino. Examples of aminocarbonylalkyl-(C₁-C₁₀) are aminocarbonylpentyl (--NHCOC₅ H₁₁) and aminocarbonylmethyl(--NHCOCH₃).

Examples of het include furan-2,4-diyl, furan-2,5-diyl, pyrrol-2,4-diyl,pyrrol-2,5-diyl, thiophen-2,4-diyl, thiophen-2,5-diyl, indol-2,5-diyl,benzofuran-2,5-diyl, benzothiophen-2,5-diyl, pyridin-2,6-diyl,pyrazin-2,6-diyl, pyrimidin-2,6-diyl, quinolin-2,6-diyl,quinoxalin-2,6-diyl, quinazolin-2,6-diyl,benzo[1,2-b:4,3-b']-dipyrrol-2,7-diyl,benzo[1,2-b:4,3-b']difuran-2,7-diyl, benzoxazol-2,5-diyl.

Examples of optionally substituted het are 3,4-dichlorofuran-2,5-diyl3,4-dimethylfuran-2,5-diyl, 3-chloropyrrol-2,5-diyl,3,4-dichloropyrrol-2,5-diyl, 3-methoxypyrol-2,5-diyl,3-methyl-4-ethylpyrrol-2,5-diyl, 3,4-difluoropyrrol-2,5-diyl,3-bromo-thiophen-2,5-diyl, 4-chloropyridin-2,6-diyl,4,5-dimethoxybenzo[1,2-b:4,3-b']-dipyrrol-2,7-diyl,3,5-dichloropyridin-2,6-diyl.

The compounds of formula B on the GENERAL FORMULA sheet can be named asderivatives of the numbering system (B') shown on the GENERAL FORMULAsheet. Such compounds will contain the1,2,3,6-tetrahydro-8-W-5-Y-benzo[1,2-b:4,3-b']dipyrrol-1-[Z--CH(X)]-structure.

The compounds of Formula I are drawn as the racemic mixture and includethe natural isomer of Formula I'a which can be resolved from the racemicmixture and/or prepared from starting materials of the natural, i.e.1(S)-configuration.

Examples of Formula I compounds of this invention include:

[S-(R*,R*)]-6,6'-[carbonylbis(5-imino-1H-indole-2-carbonyl)]bis[8-chloromethyl)-3,6,7,8-tetrahydro-1-methylbenzo[1,2-b:4,3-b']dipyrrol-4-ol(Compound 1);

[7bR-[2(7'bR*,8'aS*),7bR*,8aS*]]-2,2'-[carbonylbis(5-imino-1H-indole-2-carbonyl)]bis[1,2,8,8a-tetrahydro-7-methyl-cyclopropa(c)pyrrolo[3,2-e]indol-4(5H)-one(Compound 2);

(R*,S*)-N,N'-bis[2-[[1-(chloromethyl)-1,6-dihydro-5-hydroxy-8-methylbenzo[1,2-b:4,3-b']dipyrrol-3(2H)-yl]carbonyl]-1H-indol-5-yl]-ethanediamide(Compound 3);

[S-(R*,R*)]-6,6'-(1H-pyrrole-2,5-diyldicarbonyl)bis[8-(chloromethyl)-3,6,7,8-tertrahydro-1-methyl-benzo[1,2-b:4,3-b'-]dipyrrol-4-ol (Compound 4);

[S-(R*,R*)]-6,6'-(2,5-furandiyldicarbonyl)bis[8-(chloromethyl)-3,6,7,8-tetrahydro-1-methyl-benzo[1,2-b:4,3'b]dipyrrol-4-ol (Compound 5);

[S-(R*,R*]-N,N'-bis[2-[[1-(chloromethyl)-1,6-dihydro-5-hydroxy-8-methylbenzo[1,2-b:4,3-b']dipyrrol-3(2H)-yl]carbonyl]-1H-indol-5-yl]-2,5-furandicarboxamide(Compound 6);

[S-(R*,R*)]-N,N'-bis[2[[1-(chloromethyl)-1,6-dihydro-5-hydroxy-8-methylbenzo[1,2-b:4,3-b']dipyrrol-3(2H)-yl]carbonyl]-1H-indol-5-yl]-1H-pyrrole-2,5-dicarboxamide (Compound 7);

[R-(R*,S*)]-N,N═-bis[2-[[1-(chloromethyl)-1,6-dihydro-5-hydroxy-8-methylbenzo[1,2-b:4,3-b']dipyrrol-3(2H)-yl]carbonyl]-1H-indol-5-yl]-propanediamide (Compound 8);

[S-(R*,R*)]-6,6'-[carbonylbis(5imino-1H-indole-2-carbonyl)-bis[8-(chloromethyl)-3,6,7,8-tetrahydro-1-methyl-benzo[1,2-b:4,3b']dipyrrol-4-oldiacetate (Compound 9);

[S -(R*, R*)]-6,6'-(1H-indole-2,5-diyldicarbonyl)bis[8-(chloromethyl)-3,6,7,8-tetrahydro-1-methyl-benzo[1,2-b:4,3-b']dipyrrol-4-ol (Compound 10);

[S-(R*,R*)]-N,N'-bis[2-[[1-(chloromethyl)-1,6-dihydro-5-hydroxy-8-methylbenzo[1,2-b:4,3-b']dipyrrol-3(2H)-yl]carbonyl]-1H-indol-5-yl]-1H-indole-2,5-dicarboxamide (Compound 11 );

[S-(R*,R*)]-2-[[1-(chloromethyl)-1,6-dihydro-5-hydroxy-8-methyl-benzo[1,2-b:4,3-b']dipyrrol-3(2H)-yl]carbonyl]-N-[2-[[1-(chloromethyl)-1,6-dihydro-5-hydroxy-8-methylbenzo[1,2-b:4,3-b']dipyrrol(3(2H)-yl]carbonyl]-1H-indol-5-yl]-1H-indole-5-carboxamide (Compound 12);

[S-(R*,R*)]-5-[[1-(chloromethyl)-1,6-dihydro-5-hydroxy-8-methylbenzo[1,2-b:4,3-b']dipyrrol-3(2H)-yl]-carbonyl)-N-[2-[[1-(chloromethyl)-1,6-dihydro-5-hydroxy-8-methylbenzo[1,2-b:4,3-b']dipyrrol-3(2H)-yl]carbonyl) -1H-indol-5-yl]-1H-indole-2-carboxamide (Compound 13);

[S-(R*,R*)]-carbonylbis[imino-1H-indole-5,2-diylcarbonyl[1-(chloromethyl)-1,6-dihydro-8-methylbenzo[1,2-b:4,3-b']dipyrrole-3,5(2H) -diyl]]ester, 2,2-dimethylpropanoic acid(Compound 14);

[S-(R*,R*)]-carbonylbis[imino-1H-indole-5,2-diylcarbonyl[1-(chloromethyl)-1,6-dihydro-8-methylbenzo[1,2-b:4,3-b']dipyrrole-3,5(2H)-diyl]]ester,decanoic acid (Compound 15);

[S-(R*,R*)]-6,6'-[carbonylbis[(7,8-dihydrobenzo[1,2-b:4,3-b']dipyrrole-6,2(3H)-diyl)carbonyl]]bis[8-(chloromethyl)-3,6,7,8-tetrahydro-1-methyl-benzo[1,2-b:4,3b']dipyrrol-4-ol(Compound 16).

The compounds of Formula I are readily prepared by reacting theappropriate 4,5,8,8a-tetrahydro-4-oxocyclopropan(c)pyrrolo(3,2-e)indoleanalog (Formula B) with the biscarboxylic acid, bisisocyanate, bis-aminocarboxylate or oxycarboxylate reagent and then with the otherappropriate 4,5,8,8a-tetrahydro-4-oxocyclopropan(c)pyrrolo(3,2-e)indoleanalog (Formula B) as illustrated in Scheme 1, 2, 2a, 3, 4, 5 and 6using the following reaction conditions:

Steps 1 and 4: Run in solution of 1 mg to 100 mg/ml in solvents such asethyl acetate, methylene chloride or 1,2-dichloroethane with a gaseousacid at 1 to 5M such as HCl, or HBr or a strong organic acid such as CF₃CO₃ H. The reaction run at 0° to 50° C. The product as an amine saltisolated by evaporation of solvent and acid. The reaction may be runfrom 10 min to 48 hr.

Step 2: Reaction run in polar solvent such as DMF, DMA, or THF at -10°to 50° C. Ratio of CPI moiety: biscarboxylic acid:dehydrating agentpreferably 2:1:2. Dehydrating or coupling reagent can be a carbodiimidesuch as dicyclohexylcarbodiimide (DCC) orethyldimethylaminopropylcarbodiimide (EDC) or other amide dehydratingagents known in the art. The reaction may be run from 10 min to 72 hrs.

Steps 5 and 7: Same as Step 2 but ratio of CPI moiety:acid:dehydratingreagent is preferably 1:1:1.

Steps 3, 8, 9, 11, 14, 15 and 17: The reaction is run in aqueous organicbase. The preferred conditions are acetonitrile:water:triethylamine in aratio of 1:1:1. The ratio of these reagents may change to 1:2:2 to5:1:1. Also the solvents may change so that in place of acetonitrile maybe used DMF, DMA, N-methylpyrrolidone, THF, etc. Also the organic basecan be varied to other tertiary organic amines such as 1,5diaza-bicyclo[3.3.3)octane (DABCO), or ethyl diisopropylamine. Thetemperature is usually 0° C. to 50° C. and the reaction is usually runfor 5 min to 10 hr.

Step 6: The deprotection depends on the protecting group R. If it ist-butyl then the group may be removed by acid treatment similar toStep 1. If R=benzyl it may be removed by hydrogenolysis with Pdcatalysis in ethyl acetate or THF, for example, or by ammonium formate,methanol, water and Pd/C. If R=SiR₃ then it may be removed by acid orfluoride ions. These methods are all well known in the art.

Steps 5a and 7a: The reactions may be run in aprotic polar solvents.Preferable are pyridine, DMF, DMA, THF, or methylene chloride. When thesolvent is other than pyridine a base such as triethylamine may beadded. The reaction is run in a ratio of CPI moiety to activatedcarbonyl compound of about 1:1. The reaction may be run at -50° to 100°C. and for from 5 min to 72 hr.

Steps 10 and 16: The reagents are combined in a ratio of CPI moiety toactive carbonyl of about 2:1. The reaction may be run in polar aproticsolvents such as THF, DMF, DMA, pyridine, acetonitrile, etc. Thereaction may be run at -50° C. to 100° C. and for 10 min to severalweeks.

Steps 12, 13, 18 and 19: Ratio of CPI moiety to active carbonyl moietyof about 1:1. Otherwise run as Step 10.

In the following examples NMR spectral data were recorded in DMSO-d6unless otherwise noted. IR spectra were recorded from Nujol mulls.Solutions for UV spectra were prepared by dissolving the solid in a fewdrops of DMF or dimethylacetamide (DMA) and diluting with 95% ethanol ormethanol unless stated otherwise. TLC data is given for AnaltechUniplates of silica gel GF.

EXAMPLE 1 Preparation of[S-(R*,R*)]-6,6'-[carbonylbis(5-imino-1H-indole-2-carbonyl)]bis[8-chloromethyl)-3,6,7,8-tetrahydro-1-methyl-benzo[1,2-b:4,3-b']dipyrrol-4-ol(Compound 1)

Part A-Preparation of 5,5'-(carbonyldiimino)bis-1H-indole-2-carboxylicacid diethyl ester.

A solution of 5-aminoindole-2-carboxylic acid ethyl ester (6.82 g, 33.4mM) and 4-dimethylaminopyridine (19 mg, 0.15 mM) dissolved in THF (80mL) are cooled to -98° C. in a frozen methanol bath. To this is addeddiisopropylethylamine (7 mL, 40.2 mM) followed by 1.93M phosgene intoluene (8.5 mL, 16.4 mM). The reaction is stirred at -98° C. for 1hour, then at -78° C. for 1 hour. It is then stored at -13° C.overnight. The reaction is then quenched with water, and diluted withwater and ethyl acetate. The aqueous layer is separated and re-extractedtwice with ethyl acetate. The combined ethyl acetate layers are washedwith N HCl, giving a mixture which is separated by filtration. Thefiltered solid is found to be pure product5,5'-(carbonyldiimino)bis-1H-indole-2-carboxylic acid diethyl ester(6.38 g, 88%). The filtrate is dried and concentrated leaving 1.49 g ofdark solid. This material is chromatographed over silica gel (100 g)eluted with 10% DMF in toluene. Twenty ml fractions are collected.Product is found by TLC in fractions 23-50. Concentration of thesefractions under high vacuum leaves a residue which is crystallized fromDMF-methanol. There is thus obtained an additional 0.80 g of product.

NMR: δ 1.36(t, 6H); 4.35(q, 4H); 7.10(s, 2H); 7.26(d, 2H, J=9 Hz);7.39(d, 2H, J=9 Hz); 7.88(s, 2H); 8.52(s, 2H); 11.77(bs, 2H). IR: 3330,3260, 1705, 1695, 1545, 1250 cm⁻¹ UV: .sup.λ max (THF)=338 nm (ε=7000);301 nm (ε=28000); 292 nm (ε=26000); 250 nm (ε=31000). MS(FAB) m/z435(M+H), 406, 362, 253, 204, 159, 132. TLC: R_(f) =0.30 in (10-90)DMF-toluene.

Part B-Preparation of 5,5'-(carbonyldiimino)bis-1H-indole-2-carboxylicacid.

The indole dimer diester,5,5'-(carbonyldiimino)bis-1H-indole-2-carboxylic acid diethyl ester,(7.16 g, 16.5 mM) is treated with pyridine (300 ml) and N NaOH (60 mL).The resultant mixture is stirred 24 hrs at 25° C., 7 hrs at 50°-60° C.,and additional 48 hrs at 25° C. at which point TLC indicates Thereaction to be done. The reaction is concentrated at 25° C. under highvacuum to near dryness. The residue is acidified with 3N HCl and theresulting solid collected by filtration. The filtered solid is dissolvedin DMF (100 ml) at about 100° C. The solution is then diluted withmethanol (400 ml) and water (10 ml). The solution is cooled and thecrystalline product 5,5'-(carbonyldiimino)bis-1H-indole-2-carboxylicacid is collected by filtration (5.11 g, 82%).

NMR: ε 7.07(d, 2H, J=2 Hz); 7.28(dd, 2H, J=1.8 Hz, J=9 Hz); 7.40(d, 2H,J=9 Hz); 7.89(d, 2H, J=1.8 Hz); 8.59(bs, 2HO; 11.65(d, 2H, J=1.5 Hz).IR: 3400-2300, 1680, 1530, 1225 cm⁻¹. UV: .sup.λ max=294 nm (ε=23000).MS(FAB): m/z 379(M+H), 362, 331, 253, 176, 158, 132. TLC: R_(f) =0.33 in(40-60-2) DMF-toluene-acetic acid.

Part C-Reaction of(S)-1-(chloromethyl)-1,6-dihydro-5-hydroxy-8-methyl-benzo[1,2-b:4,3-b']dipyrrole-3(2H)-carboxylicacid 1,1-dimethyl ester [(Boc)CPI phenol chloride] with hydrogenchloride followed by condensation with5,5'-(carbonyldiimino)bis-1H-indole-2-carboxylic acid.

(S)-1-(chloromethyl)-1,6-dihydro-5-hydroxy-8-methyl-benzo[1,2-b:4,3-b']dipyrrole-3(2H)-carboxylicacid 1,1-dimethyl ester [(Boc)CPI phenol chloride] (128 mg, 0.38 mM) isdissolved in ethyl acetate (6 ml) and the solution treated with afreshly prepared solution of saturated HCl in ethyl acetate (6 ml). Thereaction is stirred one hr at which time TLC shows the completedisappearance of starting material. The reaction solution isconcentrated under vacuum and returned to atmospheric pressure undernitrogen. The resulting residue is twice re-evaporated from methylenechloride to give(S)-1-(chloromethyl)-5-hydroxy-8-methyl-1,2,3,6-tetrahydro-benzo[1,2-b:4,3-b']dipyrrolehydrochloride (CPI phenol chloride hydrochloride salt). This material istreated with a solution of the diacid,5,5'-(carbonyldiimino)bis-1H-indole-2-carboxylic acid, (78 mg, 0.21 mM)in of dry dimethylacetamide (4 ml). 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC, 68 mg, 0.08 mM) is then added as asolid. After 45 min, a second quantity of EDC is added (68 mg). Stirringis continued 50 min. The reaction is precipitated with water and thesolid centrifuged. It is washed with 0.1 NHCl, 0.5% NaHCO₃ and water,centrifuging and withdrawing the aqueous each time. The residue isevaporated in vacuo at less than 20 torr over two days. The crudeproduct is adsorbed from DMF onto Celite and chromatographed on a silicagel column (15 g) eluting with 30% DMF in toluene. Fractions of 10 mlare collected.[S-(R,,R*)]-6,6'-[carbonylbis(5-imino-1H-indole-2-carbonyl)]bis[8-chloromethyl)-3,6,7,8-tetrahydro-1-methyl-benzo[1,2-b:4,3-b']dipyrrol-4-olis found in fractions 4-16 (115.1 mg of solid, 74% yield based onBocCPI). NMR, IR, UV, FAB.

NMR: δ 2.356(S,6H); 3.582(m, 2H); 3.92(m, 2H); 4.03 (m, 2H); 4.548(m,2H); 4.67(m, 2H); 7.044(s, 4H); 7.26(d, 2H); 7.42(d, 2H); 7.64(bs, 2H);7.849(s, 2H); 8.466(bs, 2H); 9.767(bs, 2H); 10.704(bs, 2H); 11.531(bs,2H). IR: 3260, 1580, 1195, cm⁻¹. UV: ⁺ max=349 nm(ε=25000); 295nm(ε=31000). MS(FAB): m/z 815(M+H), 779, 579, 236, 201, 187, 159. DNABINDING:

    Δ[Φ].sub.λ.sup.24h ˜O

TLC: R_(f) =0.44 in (30-70)DMF-toluene.

EXAMPLE 2 Preparation of[7bR-[2(7'bR*,8'aS*),7bR*,8aS*]]-2,2'-[carbonylbis(imino-1H-indole-5,2-dicarbonyl)]bis[1,2,-8,8a-tetrahydro-7-methyl-cyclopropa(c)pyrrolo[3,2-e]indol-4(5H)-one (Compound 2)

[S-(R*,R*)]-6,6'-[carbonylbis(8imino-1H-indole-2-carbonyl)]bis[8-chloromethyl)-3,6,7,8-tetrahydro-1-methyl-benzo[1,2-b:4,3-b']dipyrrol-4-ol(80 mg, 0.1 mM) is treated with acetonitrile (10 mL), water (3 mL) andtriethylamine (3 mL). The reaction is stirred under nitrogen at roomtemperature. After 40 minutes the product which has precipitated iscollected by filtration. The filtrate is diluted with water and ethylacetate and a second quantity of product collected by filtration. Thesolids are combined giving[7bR-[2(7'bR*,8'aS*),7bR*,*aS*]]-2,2'[carbonylbis(5-imino-1H-indole-2-carbonyl)]bis[1,2,8,8a-tetrahydro-7-methyl-cyclopropan(c)pyrrolo[3,2-e]indol-4(5H)-one(38 mg, 52%).

NMR: ε 1.394(m, 2H); 2.008(s+m, 8H); 3.169(m, 2H); 4.44(m, 2H); 4.522(m,2H); 6.693(s, 2H); 6.881(s, 2H); 7.120(s, 2H); 7.266(d, 2H); 7.402(d,2H); 7.872(s, 2H); 8.512(bs, 2H); 11.536(bs, 2H); 11.675(bs, 2H). IR:3250, 1560, 1375, 1255 cm⁻¹. UV: .sup.λ max=365 nm (ε-36000); 316nm(ε=31000). MS(FAB): Calc'd. for C₄₃ H₃₅ N₈ O₅ :743.2730; found743.2737. Ions at m/z 543, 368,201. DNA BINDING:

    Δ[Φ].sub.λ.sup.24h ˜O

TLC: R_(f) =0.43 in (30-70) DMF-toluene.

EXAMPLE 3 Preparation of(R*,S*)-N,N'-bis[2-[[1-(chloromethyl)-1,6-dihydro-5-hydroxy-8-methylbenzo[1,2-b:4,3-b']dipyrrol-3(2H)- yl]carbonyl]-1H-indol-5-yl]-ethanediamide (Compound 3)

Part A-Preparation of5,5'-[(1,2-dioxo-1,2-ethanediyl)diimino]bis-1H-indole-2-carboxylic aciddiethyl ester.

5-Aminoindole-2-carboxylic acid ethyl ester (190.4 mg, 0.93 mM) isdissolved in distilled THF (15 mL) under a nitrogen atmosphere, and thesolution cooled to -78° C. Diisopropylethylamine (175 μl, 1.00 mM) issyringed in, followed by oxalyl chloride (41 μL, 0.47 mM). The reactionis stirred at -78° C. for one hr. The reaction is evaporated to halfvolume in vacuo, then treated with water. The product,5,5'-[(1,2-dioxo-1,2-ethanediyl)diimino]bis-1H-indole-2-carboxylic aciddiethyl ester, precipitates as a white solid which is collected bycentifugation and dried 72 hrs at less than 20 torr (232 mg).

NMR: δ 1.348(t, 6H); 4.35(q, 4H); 7.16(s, 2H); 7.45(d, 2H); 7.68(d, 2H);8.224(s, 2H); 10.732(bs, 2H); 11.92(bs, 2H). IR: 3360, 3320, 1690, 1670,1525, 1255, 1215 cm⁻¹. UV: .sup.λ max=330 nm (ε=5000); 299 nm (ε=10000);MS(E1): m/z 462(M+), 417, 230, 204, 158.

Part B-Preparation of5,5'-[(1,2-dioxo-1,2-ethanediyl)diimino]bis-1H-indole-2-carboxylic acid.

5,5'-[(1,2-dioxo-1,2-ethanediyl)diimino]bis-1H-indole-2-carboxylic aciddiethyl ester (220 mg, 0.48 mM) is suspended in pyridine (9 mL) andtreated with 1N NaOH (1.5 mL). The milky mixture is heated to 60° C.under nitrogen. After 24 hrs N NaOH again added (1 mL) and the reactionheated another 3 hrs. The reaction is then neutralized by the additionof 1N HCl (2.5 mL) and concentrated to one-half volume at less than 20torr. The residue is then treated with 1N HCl (50 mL) and centrifugedgiving a residual slightly pink solid. This is is washed with water,centrifuged, and dried in the vacuum oven leaving the product5,5'-[(1,2-dioxo-1,2-ethanediyl)diimino]bis-1H-indole-2-carboxylic acid(148 mg).

NMR: κ 7.098(s, 2H); 7.42(d, 2H, J=9 Hz); 7.66(d, 2H, J=9 Hz); 8.201(s,2H); 10.709(bs, 2H); 11.80(bs, 2H). IR: 3420, 3310, 1660, 1510, 1220cm⁻¹. MS(FAB): m/z 407(M+H), 385, 331, 253, 177. TLC: R_(f) =0.31 in(40-60-2) DMF-Toluene-acetic acid.

Part C-Reaction of5,5'-[(1,2-dioxo-1,2-ethanediyl)diimino]bis-1H-indole-2-carboxylic acidwith(S)-1-(chloromethyl)-1,6-dihydro-5-hydroxy-8-methyl-benzo[1,2-b:4,3-b']dipyrrole-3(2H)carboxylicacid 1,1-dimethyl ester [(Boc)CPI phenol chloride].

(S)-1-(chloromethyl)-1,6-dihydro-5-hydroxy-8-methyl-benzo[1,2-b:4,3-b']dipyrrole-3(2H)-carboxylicacid 1,1-dimethyl ester [(Boc)CPI phenol chloride] (42.5 mg, 0.13 mM) isdissolved in ethyl acetate (1 mL) under a nitrogen atmosphere in thedark. The reaction is treated with ethyl acetate (3 mL) saturated withHCl. The reaction is stirred 1 hr and evaporated in vacuo. Nitrogen islet in when the vacuum is released.(S)-1-(chloromethyl)-5-hydroxy-8-methyl-1,2,3,6-tetrahydro-benzo[1,2-b:4,3-b']dipyrrolehydrochloride (CPI phenol chloride hydrochloride) as a solid is obtainedon re-evaporation 2× with CH₂ C;₂. A suspension of5,5'-[(1,2-dioxo-1,2-ethanediyl)diimino]bis-1H-indole-2-carboxylic acid(28.4 mg, 0.06 mM) in dimethylacetamide (2 mL) is added to the reaction.EDC is added as a solid (43.4 mg, 0.23 mM). The reaction is stirred inthe dark under nitrogen at room temperature for 100 min. Water is addedto precipitate the product. The solid, isolated by centrifugation, iswashed twice with 5% NaHCO₃, then with 0.1N HCl and water. The solid isdried under vacuum. The solid is adsorbed from DMF onto silica gel, andchromatographed on a silica gel column (3 g) in 20% to 30% DMF intoluene. Fractions of 1-2 mL are collected.(R*,S*)-N,N'-bis[2-[[1-(chloromethyl)-1,6-dihydro-5-hydroxy-8-methylbenzo[1,2-[b:4,3b']dipyrrol-3(2H)-yl]carbonyl]-1H-indol-5-yl]-ethanediamideis found in fractions 15-22 (24.6 mg, 45% yield).

NMR: (DMF-d7) δ 2.216(s, 6H); 3.47(m, 2H); 3.77(m, 2H); 3.918(m, 2H);4.51(m, 4H); 6.95(s, 2H); 7.03(s, 2H); 7.40(d, 2H); 7.62(d, 2H);8.22(s,2H). UV: .sup.λ max=341 nm (ε=38000); 293 nm (ε=52000). TLC:R_(f) =0.58 in (30-70) DMF-toluene.

EXAMPLE 4 Preparation of[S-(R*,R*)]-6,6'-(1H-pyrrole-2,5-diyldicarbonyl)bis[8-(chloromethyl)-3,6,7,8-benzo[1,2-b:4,3-b']dipyrrol-4-ol(Compound 4)

(S)-1-(chloromethyl)-1,6-dihydro-5-hydroxy-8-methyl-benzo[1,2-b:4,3-b']dipyrrole-3(2H)-carboxylicacid 1,1-dimethyl ester [(Boc)CPI phenol chloride] (40 mg, 0.12 nM) isdissolved in ethyl acetate (1 mL) under a nitrogen atmosphere. Thereaction is treated with ethyl acetate freshly saturated with HCl (3 ml)and the resultant solution stirred at room temperature for 1 hr. Thesolvent is removed in vacuo, and the(S)-1-(chloromethyl)-5-hydroxy-8-methyl-1,2,3,6-tetra-hydro-benzo[1,2-b:4,3-b']dipyrrolehydrochloride (CPI phenol chloride hydrochloride) as a purplish residualsolid is re-evaporated 2× with CH₂ CL₂, flushing with nitrogen betweenevaporations. This is treated with pyrrole-2,5-dicarboxylic acid (8.9mg, 0.057 mM) in dry dimethylacetamide (1 mL) and EDC (24 mg, 0.125 mM)and the resultant solution stirred at 25 C. After 50 min. and additionalquantity of EDC is added (24 mg, 0.125 mM). After an additonal 55 minthe reaction is diluted with water (5 mL) and saturated aqueous sodiumchloride solution (5 mL) and extracted with ethyl acetate 4 times. Thecombined organic layers are dried (Na₂ SO₄) and evaporated. The cruderesidue is adsorbed from ethyl acetate-acetone onto 0.5 g silica gel andchromatographed on a 6 g silica gel column in 10% DMF-toluene followedby 15% DMF-toluene. Fractions of 3-4 ml are collected.[S(R*,R*)]-6,6'-(1H-pyrrole-2,5-diyldicarbonyl)bis[8-(chloromethyl)-3,6,7,8-benzo[1,2-b:4,3-b'-]dipyrrol-4-olis found in Fr. 23-37 (25 mg).

NMR: δ 2.349(s, 6H); 3.59(m, 2H); 3.90(m, 2H); 4.01(m, 2H); 4.40(m, 2H);4.55(m, 2H); 6.850(s, 2H); 7.039(m, 2H); 7.60(m, 2H); 9.77(s, 2H);10.71(bs, 2H);11.36(bs, 1H). UV: .sup.λ max=356 nm (ε=16000); 290 nm(ε=21000). TLC: R_(f) =0.47 in (20-80) DMF-toluene.

EXAMPLE 5 Preparation of[S-(R*,R*)]-6,6'-(2,5-furandiyldicarbonyl)bis[8-(chloromethyl)-3,6,7,8-tetrahydro-1-methyl-benzo[1,2-b4,3-b']dipyrrol-4-ol (Compound 5).

(S)-1-(chloromethyl)-1,6-dihydro-5-hydroxy-8-methyl-benzo[1,2-b:4,3-b']dipyrrole-3(2H)-carboxylicacid 1,1-dimethyl ester [(Boc)CPI phenol chloride] (40.6 mg, 0.12 mM) isdissolved in ethyl acetate (1 mL), put under nitrogen, and treated withHCl-saturated ethyl acetate (3 mL). After 50 minutes, the reaction isevaporated in vacuo, returning to atmospheric pressure under nitrogen.(S)-1-(chloromethyl)-5-hydroxy-8-methyl-1,2,3,6-tetrahydro-benzo[1,2-b:4,3-b']dipyrrolehydrochloride (CPI phenol chloride hydrochloride) as a solid is obtainedon re-evaporation 2× from CH₂ Cl₂. The solid is treated with a solutionof furan-2,5-dicarboxylic acid (9.7 mg, 0.062 mM) in dry dimethylacetamide (1 mL). EDC (43.1 mg 0.22 mM) is added as a solid in 2 equalportions, 45 min apart. The reaction is stopped 170 min after the secondaddition, by adding ethyl acetate and water. The water is re-extractedwith ethyl acetate. The combined organic layers are washed with 0.5NHCl, 1% NaHCO₃, and water. Drying (Na₂ SO₄) and concentration gives ayellow solid (55.2 mg). The solid is adsorbed from DMF-acetone ontosilica gel (0.7 g). This is placed on a silica gel column (7 g), elutingwith 10% to 15% DMF in toluene. Fractions of 3-4 mL are collected.[S-(R*,R*)]-6,6'-(2,5-furandiyl-dicarbonyl)bis[8-(chloromethyl)-3,6,7,8-tetrahydro-1-methyl-benzo[1,2-b4,3-b']dipyrrol-4-ol, as a bright yellow solid, is isolated byconcentrating fractions 23-31 (22.3 mg, 63% yield).

NMR: δ 2.344(s, 6H); 3.63(m, 2H); 3.89(m, 2H); 4.029(m, 2H); 4.51(m,2H); 4.60(m, 2H); 7.048(s, 2H); 7.37(s, 2H); 7.64(m, 2H); 9.82(s, 2H);10.75(bs, 2H). UV: .sup.λ max=360 nm (ε=20000); 295 nm (ε=24000). TLC:R_(f) =0.47 in (20-80) DMF-toluene.

EXAMPLE 6 Preparation of[S-(R*,R*]-N,N'-bis[2-[[1-(chloro-methyl)-1,6-dihydro-5-hydroxy-8-methylbenzo[1,2-b:4,3-b']dipyrrol-3(2H)-yl]carbonyl]-1H-indol-5-yl]-2,5-furandicarboxamide(Compound 6)

Part A-Preparation of5,5'-[2,5-furandiylbis(carbonylimino)]-bis-1H-indole-2-carboxylic aciddiethyl ester.

Furan-2,5-dicarboxylic acid (50 mg, 0.32 mM, A Sohst and B. Tollens,Justig Liebigs Ann. Chem., 245, 1 (1888)) and5-amino-indole-2-carboxylic acid ethyl ester (130 mg, 0.64 mM) aredissolved in dimethylacetamide (2 mL) and the solution treated with EDC(132 mg, 0.69 mM). After 20 hrs the reaction is diluted with water (20mL) and extracted twice with ethyl acetate. The combined ethyl acetatesolutions are washed with N HCl and N NaOH, and dried (MgSO₄).Concentration of the ethyl acetate in vacuo leaves a residue (154 mg).This is chromatographed over 10 g of silica gel eluting with 50% to 60%ethyl acetate in hexane followed by 50% ethyl acetate in toluene.Fractions of 4 mL are collected.5,5'-[2,5-furandiylbis(carbonylimino)]bis-1H-indole-2-carboxylic aciddiethyl ester (107 mg) is obtained on evaporation of fr 22-50.

NMR: δ 1.413(t, 6H); 4.38(g, 4H); 7.137(s, 2H); 7.51(d, 2H); 7.58(d,2H); 8.021(s, 2H); 8.097(bs, 2H); 10.146(bs, 2H); 11.742(bs, 2H). TLC:R_(f) =0.38 in (60-40) ethyl acetate-hexane.

Part B-Preparation of5,5'-[2,5-furandiylbis(carbonylimino)]-bis-1H-indole-2-carboxylic acid.

5,5'-[2,5-furandiylbis(carbonylimino)]bis-1H-indole-2-carboxylic aciddiethyl ester (60 mg, 0.11 mM) is suspended in methanol (0.4 mL) andtreated with N NaOH (0.25 mL). After 5 days the reaction is evaporatedin vacuo and the residue dissovlved in water and the solution acidifiedto pH 2 with N HCl. This solution is freeze dried leaving5,5'-[2,5-furandiylbis(carbonylimino)]bis-1H-indole-2-carboxylic acid(22 mg).

Part C-Reaction of5,5'-[2,5-furandiylbis(carbonylimino)]bis-1H-indole-2-carboxylic acidwith (Boc)CPI phenol chloride.

(S)-1-(chloromethyl)-1,6-dihydro-5-hydroxy-8-methyl-benzo[1,2-b:4,3-b']dipyrrole-3(2H)-carboxylicacid 1,1-dimethyl ester [(Boc)CPI phenol chloride] (33 mg, 0.098 mM) isdissolved in ethyl acetate (1 mL) and the resultant solution stirredunder an Ar atmosphere and treated with a freshly prepared saturated HClsolution in ethyl acetate (3 mL). After 45 min the mixture is evaporatedin vacuo and the resultant(S)-1-(chloromethyl)-5-hydroxy-8-methyl-1,2,3,6-tetrahydro-benzo[1,2-b:4,3-b']dipyrrolehydrochloride (CPI phenol chloride hydrochloride) twice treated with CH₂Cl₂ and re-evaporated. This is treated with dimethylacetamide (1 mL)containing5,5'-[2,5-furandiylbis(carbonylimino]bis-1H-indole-2-carboxylic acid (22mg, 0.047 mg). The resultant solution is treated in 2 portions about 50min apart with EDC (40 mg, 0.21 mM). After 50 min the reaction isdiluted with water and mixture extracted with THF-ethyl acetate. Thecombined organic layers are dried over MgSO₄ and concentrated in vacuoleaving a solid (15 mg). This material is chromatographed over silicagel (5 g) eluted with 20% to 30 % DMF in toluene. Fractions of 3 mL arecollected.[S-(R*,R*]-N,N'-bis[2-[[1-(chloromethyl)-1,6-dihydro-5-hydroxy-8-methylbenzo[1,2-b:4,3-b']dipyrrol-3(2H)-yl]-carbonyl]-1H-indol-5-yl]-2,5-furandicarboxamide(5 mg) is obtained on evaporation of fr 16-28.

NMR: δ 2.363(s, 6H); 3.60(m, 2H); 3.90(m, 2H); 4.03(m, 2H); 4.56(m, 2H);4.68(m, 2H); 6.616(s, 1H); 7.05(s, 2H); 7.16(m, 2H); 7.41(s, 2H);7.53(s, 4H); 7.64(m, 1H); 8.16(s, 2H); 9.79(s, 2H); 10.29(bs, 2H);10.73(bs, 2H); 11.73(bs, 2H). UV: .sup.λ max (MeOH)=350 nm (ε=41000);283 nm (ε=50500). MS(FAB): m/z 911 (M+H), 909 (M+H), 412. 395, 335, 300,253, 210. TLC: R_(f) =0.217 in (20-80) DMF-toluene.

EXAMPLE 7 Preparation of[S-(R*,R*)]-N,N'-bis[2[[1-(chloromethyl)-1,6-dihydro-5-hydroxy-8-methylbenzo[1,2-b4,3 -b']dipyrrol-3(2H) -yl]carbonyl]-1H-indol-5-yl]-1H-pyrrole-2,5-dicarboxamide (Compound 7)

Part A-Coupling of pyrrole-2,5-dicarboxylic acid to5-amino-indole-2-carboxylic acid ethyl ester.

Pyrrole-2,5-dicarboxylic acid (50 mg, 0. 32 mM, Kuhn, R. and Dury, K.,Justus Liebigs. Ann., Chem,. 571, 44 (1951)) and5-amino-indole-2-carboxylic acid ethyl ester (130 mg, 0.64 mM) aredissolved in dimethylacetamide (2 mL) and the solution treated with EDC(129 mg, 0.67 mM). After 22 hrs the mixture is treated with ice water(20 mL) and the precipitated solid filtered. This material ischromatographed over silica gel (10 g), eluted with 10% to 15% DMF intoluene. Fractions of 4 mL are collected. A residue containing theproduct (72 mg) is obtained on evaporation of fr. 22-42. This residue istriturated with ethyl acetate and dried leaving5,5'-[1H-pyrrole-2,5-diylbis(carbonylimino)-bis-1H-indole-2-carboxylicacid diethyl ester (42 mg).

NMR: δ 1.413(t, 6H); 4:38(q, 4H); 7.137(s, 2H); 7.51(d, 2H); 7.58(d,2H); 8.021 (s, 2H); 8.097(bs, 2H); 10.146(bs, 2H); 11,742(bs, 2H). TLC:R_(f) =0.38 in (60-40) ethyl acetate-hexane.

Part B-Preparation of5,5'-[1H-pyrrole-2,5-diylbis(carbonylimino)-bis-1H-indole-2-carboxylicacid

5,5'-[1H-pyrrole-2,5-diylbis(carbonylimino)-bis-1H-indole-2-carboxylicacid diethyl ester (42 mg, 08 mM) is dissolved in pyridine (1 mL) andthe solution treated with N NaOH (0.2 mL). After 24 hrs the reation istreated with additional N NaOH (0.4 mL). After an additional 96 hrs thereaction is evaporated in vacuo and the residue dissolved in water (5mL). The aqueous solution is acidified with N HCl to pH 2 and thenfreeze-dried, leaving5,5'-[1H-pyrrole-2,5-diylbis(carbonylimino)-bis-1H-indole-2-carboxylicacid.

TLC: R_(f) =0.18 in (2-15-85) acetic acid-DMF-toluene.

Part C-Coupling of5,5'-[1H-pyrrole-2,5-diylbis(carbonylimino)-bis-1H-indole-2-carboxylicacid with(S)-1-(chloromethyl)-1,6-dihydro-5-hydroxy-8-methyl-benzo[1,2-b:4,3-b']dipyrrole-3(2H)-carboxylicacid 1,1-dimethyl ester [(Boc)CPI phenol chloride]

(S)-1-(chloromethyl)-1,6-dihydro-5-hydroxy-8-methyl-benzo[1,2-b:4,3-b']dipyrrole-3(2H)-carboxylic acid 1,1-dimethyl ester [(Boc)CPI phenol chloride] (40 mg,0.12 nM) is dissolved in ethyl acetate (1 mL) under a nitrogenatmosphere. The reaction is treated with ethyl acetate freshly saturatedwith HCl (3 mL) and the resultant solution stirred at room temperaturefor 45 min. The solvent is removed in vacuo, and the residue returned toatmospheric pressure with nitrogen. The residue is twice re-evaporatedwith CH₂ Cl₂ leaving(S)-1-(chloromethyl)-5-hydroxy-8-methyl-1,2,3,6-tetrahydro-benzo[1,2-b:4,3-b']dipyrrolehydrochloride (CPI phenol chloride hydrochloride). This material isdissolved in dimethylacetamide (1 mL) and the solution treated with5,5'-[1H-pyrrole-2,5-diylbis(carbonylimino)-bis-1H-indole-2-carboxylicacid (28 mg) and with EDC (25 mg, 0.13 mM). After 50 min the reaction istreated with additional EDC (25 mg). After another 50 min the reactionis diluted with water and the precipitated solids partitioned intoTHF-ethyl acetate. The combined organic layers are dried (MgSO₄) andconcentrated in vacuo leaving a solid (80 mg). This material ischromatographed over of silica gel (10 g) eluted with 20% DMF intoluene. Fractions of 3 mL are collected.[S-(R*,R*)]-N,N'-bis[2[[1-(chloromethyl)-1,6-dihydro-5-hydroxy-8-methylbenzo[1,2-b4,3-b']dipyrrol-3(2H)-yl]carbonyl]-1H-indol-5-yl]-1H-pyrrole-2,5-dicarboxamide(11 mg) is obtained on evaporation of fr 26-45.

NMR: δ 2.363(s, 6H); 3.60(m, 2H); 3.91(m, 2H); 4.032(m, 2H); 4.54(m,2H); 4.68(m, 2H); 6.625(s, 1H); 7.05(s, 2H); 7.09(s, 2H); 7.13(s, 2H);7.49(m, 4H); 7.64(m, 1H); 8.18(s, 2H); 9.78(s, 2H); 10.09 (bs, 2H);10.72(bs, 2H); 11.67(bs, 2H); 12.22(bs, 1H). UV: .sup.λmax(methanol)=344 nm (ε=58600); 296 nm (ε=68000). MS(FAB): Calc'd forC₄₈ H₃₉ Cl₂ N₉ O₆ ; 908.2478; Found: 908.2496. ions at m/z 543, 368,201. TLC: R_(f) =0.21 in (20-80)DMF-toluene.

EXAMPLE 8 Preparation of[R-(R*,S*)]-N,N'-bis[2-[[1-(chloro-methyl)-1,6-dihydro-5-hydroxy-8-methylbenzo[1,2-b4,3-b']dipyrrol-3(2H)-yl]carbonyl]-1H-indol-5-yl]-propane-diamide(Compound 8)

Part A-Reaction of 5-aminoindole-2-carboxylic acid ethyl ester withmalonyl dichloride.

A solution of 5-aminoindole-2-carboxylic acid ethyl ester (196 mg, 0.96mM) in dry distilled THF (15 ml) is cooled in an ice bath undernitrogen. N,N-Diisopropylethylamine (350 μL, 2.01 mM) and malonyldichloride (80 μL, 0.82 mM) are added by syringe. After one hour thesolvent is partially removed and5,5'-[(1,3-dioxo-1,3-propanediyl)-diimino]bis-1H-indole-2-carboxylicacid diethyl ester precipitated with water. The mixture is centrifuged,and the liquid layer withdrawn. The solid is dried under vacuum. Theresidue is chromatographed over silica gel (2 g) eluted with 25% acetonein toluene, 40% acetone in toluene, 60% acetone in toluene, 80% acetonein toluene and acetone. Fractions of 15 mL are collected. The product5,5'-[(1,3-dioxo-1,3-propanediyl)-diimino]bis-1H-indole-2-carboxylicacid diethyl ester is found in fr 9-13 (77 mg).

NMR: (Acetone-d6) δ 1.37(t, 6H); 3.571(s, 2H); 4.36(q, 4H); 7.16(s, 2H);7.48(m, 4H); 8.17(s, 2H); 9.71(bs, 2H); 10.92(bs, 2H). IR: 3300, 1650,1200 cm⁻¹. UV: .sup.λ max=340 nm (ε=5400); 297 nm (ε=28000). MS(E1):m/z476(M+), 340, 272, 226, 204, 158. TLC: R_(f) =0.21 in (40-60)Acetone-hexane.

Part B-Preparation of5,5'-[(1,3-dioxo-1,3-propanediyl)diimino]bis-1H-indole-2-carboxylicacid.

The diester of Part A (0.16 mM) is dissolved in pyridine (3 ml) and 1Nsodium Hydroxide (0.53 ml). The reaction mixture is stirred undernitrogen at 50° C. for about 2 hours, then at room temperature for 4hours. The reaction mixture is treated with 1N HCl and evaporated tonear dryness. The product is precipitated with water and isolated bycentrifugation, and washed with water, leaving5,5'-[(1,3-dioxo-1,3-propanediyl)-diimino]bis-1H-indole-2-carboxylicacid.

NMR: δ 3.47(s,2H); 7.04(s,2H); 7.36(m,4H); 8.03(s,2H); 10.09(bs,2H);11.68(bs,2H). UV: .sup.λ max=294 nm (ε=13000).

Part C-Coupling of5,5'-[(1,3-dioxo-1,3-propanediyl)diimino]-bis-1H-indole-2-carboxylicacid and(S)-1-(chloromethyl)-1,6-dihydro-5-hydroxy-8-methyl-benzo[1,2-b:4,3-b']dipyrrole-3(2H)-carboxylicacid 1,1-dimethyl ester [(Boc)CPI phenol chloride].

(S)-1-(chloromethyl)-1,6-dihydro-5-hydroxy-8-methyl-benzo[1,2-b:4,3-b']dipyrrole-3(2H)-carboxylicacid 1,1-dimethyl ester [(Boc)CPI phenol chloride] (37.8 mg, 0.11 mM) isdissolved in ethyl acetate (0.5 mL) under a nitrogen atmosphere. Thesolution is treated with HCl-saturated ethyl acetate (3 mL) for one hr.The reaction is evaporated, releasing the vacuum with nitrogen. Theresultant(S)-1-(chloromethyl)-5-hydroxy-8-methyl-1,2,3,6-tetrahydro-benzo[1,2-b:4,3-b']dipyrrolehydrochloride (CPI phenol chloride hydrochloride) is re-evaporated 2×with CH₂ Cl₂. This material is treated with5,5'-[(1,3-dioxo-1,3-propanediyl)diimino]bis-1H-indole-2-carboxylicacid, (24.3 mg, 0.058 mM) in dry dimethylacetamide (1 mL). EDC (49 mg,0.156 mM) is weighed out and about one half is added to the reaction.The mixture is stirred at room temperature 45 min. Then the rest of theEDC is added, and stirring is continued 40 min. The reaction is dilutedwith ethyl acetate and water. The insoluble materials are removed byfiltration and saved. The filtrate is separated into layers and theaqueous re-extrated with ethyl acetate. The combined organic layers aredried (Na₂ SO₄) and concentrated in vacuum. The residue is combined withthe filtered solids from above and are absorbed onto silica gel (0.4 g)and added to the top of a silica gel column (5 g). The column is elutedwith 22% to 28% DMF in toluene, collecting fractions of 2 mL. Theproduct5,5'-[(1,3-dioxo-1,3-propanediyl)diimino]bis-1H-indole-2-carboxylic acidis isolated in fractions 14-28 (36 mg).

NMR: δ 2.354(s, 6H); 3.51(m, 2H); 3.60(m, 2H); 3.91(m, 2H); 4.03(m, 2H);4.52(m, 2H); 4.67(m, 2H); 7.05(s, 2H); 7.10(s, 2H); 7.38(d, 2H); 7.46(d,2H); 7.64(m, 2H); 8.10(s, 2H); 9.78(s, 2H); 10.13(bs, 2H); 10.72(bs,2H); 11.64(bs, 2H). UV: .sup.λ max=336 nm(ε=35000); 294 nm (ε=44000).TLC: R_(f) =0.46 in (30-70) DMF-toluene.

EXAMPLE 9 Preparation of[S-(R*,R*)]-6,6'-[carbonylbis(5-imino-1H-indole-2-carbonyl)]bis[8-(chloromethyl)-3,6,7,8-tetrahydro-1-methyl-benzo[1,2-b:4,3-b']dipyrrol-4-oldiacetate (Compound 9).

[7bR-[2(7'bR*,8'aS*),7bR*,8aS,]]-2,2'-[carbonylbis(imino-1H-indole-5,2-dicarbonyl)]bis[1,2,8,8a-tetrahydro-7-methyl-cyclopropa(c)pyrrolo[3,2-e]indol-4(5H)-one(5 mg, 0.007 mM) is dissolved in DMF (200 μl) and acetone (0.5 ml) andthe solution treated with pyridine hydrochloride (8.3 mg, 0.07 mM). Thereaction is stirred at room temperature in the dark for 110 minutes andthen concentrated to a small volume, and treated with water. The mixtureis centrifuged and the liquids removed leaving[S-(R*,R*)]-6,6'-[carbonylbis(imino-1H-indole-5,2-dicarbonyl)]bis[8-chloromethyl)-3,6,7,8-tetrahydro-1-methyl-benzo[1,2-b:4,3-b']dipyrrol-4-olas a solid. The solid is dissolved in pyridine (100 μL) and treated withacetic anhydride (6 μL, 0.06 mM). The reaction is stirred in the dark atroom temperature for 85 minutes, quenched with water and concentrated todryness. The residue is treated with water and centrifuged. The liquidis removed and the solid dried under vacuum. This solid is adsorbed fromDMF onto silica gel (0.07 g). Silica with absorbed compound is placed ona silica gel column (1 g), eluting with 10% to 40% DMF in toluene.Fractions of 1/2 mL are taken.[S-(R*,R*)]-6,6'-[carbonyl-bis(5-imino-1H-indole-2-carbonyl)]bis[8-(chloromethyl)-3,6,7,8-tetrahydro-1-methyl-benzo[1,2-b4,3-b']dipyrrol-4-ol diacetate elutes in fractions 38-47 (3.4 mg).

NMR(DMSO-d6, TMS):δ 2.37 (s, 6H); 2.40(s, 6H); 3.72(m, 2H); 3.99 (m,2H); 4.19 (m, 2H); 4.61(m, 2H); 4.74(m, 2H); 7.09 (s, 2H); 7.24 (m, 4H);7.43 (m, 2H); 7.83 (bs, 2H); 7.87 (m, 2H); 8.50 (bs, 2H); 11.12 (bs,2H); 11.56 (bs, 2H). UV(DMA+MeOH): λmax=325 nm (ε=43,800); 294 nm(ε=43,800).

EXAMPLE 10 Preparation of[S-(R*,R*)]-6,6'-(1H-indole-2,5-diyl-dicarbonyl)bis[8-(chloromethyl)-3,6,7,8-tetrahydro-1-methyl-benzo[1,2-b:4,3-b']dipyrrol-4-ol(Compound 10)

(S)-1-(chloromethyl)-1,6-dihydro-5-hydroxy-8-methyl-benzo[1,2-b:4,3-b']dipyrrole-3(2H)-carboxylicacid 1,1-dimethyl ester [(Boc)CPI phenol chloride] (49 mg, 0.146 mM) isstirred at room temperature under nitrogen in ethyl acetate (0.6 mL).Ethyl acetate saturated with gaseous HCl (3 mL) is added and thereaction followed by TLC and is complete in about 45 minutes. Thereaction is evaporated, releasing the vacuum with nitrogen. Theresultant(S)-1-(chloromethyl)-5-hydroxy-8-methyl-1,2,3,6-tetrahydro-benzo[1,2-b:4,3-b']dipyrrolehydrochloride (CPI phenol chloride hydrochloride) is re-evaporated 2×with CH₂ Cl₂. The residual CPI phenol chloride hydrochloride salt isdissolved in dry dimethylacetamide (1 mL) and stirred at roomtemperature under nitrogen. Indole-2,5-dicarboxylic acid (15 mg, 0.073mM) is added, followed by EDC (40 mg, 0.21 mM). After 45 minutes,additional EDC (23 mg, 0.12 mM) is added to the reaction mixture andwhich is then left to react for 1 hour. The reaction mixture istransferred to a centrifuge tube, rinsing in reaction with DMF (0.5 mL)and diluted with water to precipitate the product which is centrifugedand decanted. The crude product is transferred to a round bottom flaskwith acetone and the resulting solution evaporated under vacuum. Thecrude product is coated on silica gel (0.5 g) and chromatographed over asilica gel column (6 g) made up in 20% DMF in toluene and eluted withthe same solvent collecting 1 ml fractions.[S-(R*,R*)]-6,6'-(1H-indole-2,5-diyldicarbonyl)bis[8-(chloromethyl)-3,6,7,8-tetrahydro-1-methyl-benzo[1,2-b:4,3-b']dipyrrol-4-olis collected in fractions 22-47 and obtained on evaporation to dryness(31 mg).

NMR(d₆ -DMSO,TMS): δ 2.33(s, 3H); 2.36(s, 3H); 3.54-3.68(t, 2H);3.79-3.96(m, 3H); 3.98-4.11(t, 2H); 4.23-4.44(bs, 1H); 4.50-4.60(d, 1H);4.62-4.75(t, 1H); 7.02(s, 1H); 7.05(s, 1H); 7.22(s, 1H); 7.44-7.53(d,1H); 7.53-7.62(d, 1H); 7.62-7.73(bs, 1H); 7.95(s, 1H); 7.98(s, 1H);9.60-9.75(bs, 1H); 9.80(s, 1H); 10.66(s, 1H); 10.73(s, 1H); 11.93(s,1H).

EXAMPLE 11 Preparation of[S-(R*,R*)]-N,N'-bis[2-[[1-(chloromethyl)-1,6-dihydro-5-hydroxy-8-methylbenzo[1,2-b:4,3-b']dipyrrol-3(2H)-yl]carbonyl]-1H-indol-5-yl]-1H-indole-2,5-dicarboxamide(Compound 11).

Part A-Preparation of5,5'-[1H-indole-2,5-diylbis(carbonyl-imino)bis-1H-indole-2-carboxylicacid diethyl ester.

Indole-2,5-dicarboxylic acid (200 mg, 0.49 mM) is stirred at roomtemperature under nitrogen in dry DMF (4 mL). To this is added5-amino-indole-2-carboxylic acid ethyl ester (400 mg, 1.96 mM) and EDC(200 mg, 1.05 mM). After 3 days, the reaction mixture is transferred toa centrifuge tube and diluted with water. The precipitated solid iscentrifuged and the supernatant liquid decanted. The residue is washedwith acetone leaving5,5'-[1H-indole-2,5-diyl-bis(carbonylimino)bis-1H-indole-2-carboxylicacid diethyl ester (78 mg). Additional5,5'-[1H-indole-2,5-diylbis(carbonylimino)bis-1H-indole-2-carboxylicacid diethyl ester is obtained from the above acetone washings byconcentration to dryness and chromatography over silica gel (40 g)eluting with 10%-20% DMF in toluene. Factions of 15 mL are collected andthe 5,5'-[1H-indole-2,5-diylbis(carbonylimino)bis-1H-indole-2-carboxylicacid diethyl ester (83 mg) is obtained on evaporation to dryness offractions 35-40.

Part B-Preparation of5,5'-[1H-indole-2,5-diylbis(carbonylimino)]bis-1H-indole-2-carboxylicacid.

5,5'-[1H-indole-2,5-diylbis(carbonylimino)]bis-1H-indole-2-carboxylicacid diethyl ester (161 mg, 0.28 mM) is dissolved with stirring undernitrogen at room temperature in pyridine (5 mL) absolute ethanol (5 mL).To the resultant solution is added N NaOH (1 mL) and which is allowed tostand for 5 hr. The solvent is then mostly evaporated under vacuum,water is added, and the reaction mixture freeze-dried. The resultantcrude product is coated on Celite (1.5 g) and added to the top of a C-18reverse phase silica gel column (15 g). The column is eluted with thefollowing: 50% DMF in water (200 mL); 60% DMF in water (100 mL); 70%DMF-30% water (100 ml); 80% DMF-20% water (300 ml). Fractions of 5 mlare collected, analyzing them by TLC.5,5'-[1H-indole-2,5-diylbis(carbonylimino) ]bis-1H-indole-2-carboxylicacid (46 mg) is found in fractions 7-21.

Part C-Reaction of5,5'-[1H-indole-2,5-diylbis(carbonylimino)]bis-1H-indole-2-carboxylicacid with (Boc)CPI phenol chloride.

(S)-1-(chloromethyl)-1,6-dihydro-5-hydroxy-8-methyl-benzo[1,2-b:4,3-b']dipyrrole-3(2H)-carboxylic acid 1,1-dimethyl ester [(Boc)CPIphenol chloride] (59 mg, 0.176 mM) is stirred at room temperature undernitrogen in dark for 45 min in ethyl acetate (1 mL) and ethyl acetatesaturated with gaseous HCl (4 mL). TLC after 30 min shows reaction to becomplete. The reaction mixture is evaporated under vacuum, andre-evaporated with CH₂ Cl₂ 2×, letting in nitrogen each time giving(S)-1-(chloromethyl)-5-hydroxy-8-methyl-1,2,3,6-tetrahydro-benzo[1,2-b:4,3-b']dipyrrolehydrochloride (CPI phenol chloride hydrochloride). The CPI phenolchloride hydrochloride salt is dissolved with stirring under nitrogen indark in dry DMA (1 mL). The solution is treated with5,5'-[1H-indole-2,5-diylbis(carbonylimino)]bis-1H-indole-2-carboxylicacid (46 mg, 0.088 mM) and EDC (50 mg, 0.26 mM). After 30 min additionalEDC is added (26 mg, 0.13 mM). The reaction is allowed to stand anadditional 1 hr then transferred to a centrifuge tube and washed in withDMF (1 mL). Water is added to precipitate product which is thencollected by centrifugation. The solid is washed into a RB flask withacetone and evaporated under vacuum. The residue is coated on silica gel(1 g) and chromatographed over a silica gel column (9 g) . The column iseluted with 25% DMF in toluene followed by 30% and 40% DMF in toluene.Fractions of 2 ml are collected, and analyzed by TLC.[S-(R*,R*)]-N,N'-bis[2-[[1-(chloromethyl)-1,6-dihydro-5-hydroxy-8-methylbenzo-[1,2-b4,3-b']dipyrrol-3(2H)-yl]carbonyl]-1H-indol-5-yl]-1H-indole-2,5-dicarboxamideis found in fractions 31-80. Impure product is found in fractions 8-30.These are rechromatographed over a silica gel column (4.5 g) eluted with20% DMf in toluene to 30% DMF in toluene. Fractions of 1 mL arecollected.[S-(R*,R*)]-N,N'-bis[2-[[1-(chloro-methyl)-1,6-dihydro-5-hydroxy-8-methylbenzo[1,2-b4,3-b']dipyrrol-3(2H)-yl]carbonyl]-1H-indol-5-yl]-1H-indole-2,5-dicarboxamideis found in fractions 25-75. Combination of the material from the twochromatographies gives[S-(R*,R*)]-N,N'-bis[2-[[1-(chloromethyl)-1,6-dihydro-5-hydroxy-8-methylbenzo[1,2-b4,3-b']dipyrrol-3(2H)-yl]carbonyl]-1H-indol-5-yl]-1H-indole-2,5-dicarboxamide(47 mg).

NMR(d₆ -DMSO,TMS): δ 2.37(s, 6H); 3.55-3.68(t, 2H); 3.87-3.97(d, 2H);3.97-4.10(t, 2H); 4.51-4.61(d, 2H); 4.61-4.77(t, 2H); 7.07(s, 2H);7.14(s, 1H); 7.17(s, 1H); 7.45-7.75(m, 8H); 7.96(s, 1H); 8.23(s, 2H);8.26(s, 1H); 8.46(s, 1H); 9.82(s, 2H); 10.18(s, 1H); 10.33(s, 1H);10.75(s, 2H); 11.66(s, 1H); 11.71(s, 1H); 12.09(s, 1H).

UV(DMA+MeOH): λmax=328 nm (ε=33,560); 281 nm (ε=38,350)

EXAMPLE 12 Preparation of[S-(R*,R*)]-2-[[1-(chloromethyl)-1,6-dihydro-5-hydroxy-8-methyl-benzo[1,2-b:4,3-b']dipyrrol-3(2H)-yl]-carbonyl]-N-[2-[[1-(chloromethyl)-1,6-dihydro-5-hydroxy-8-methylbenzo[1,2-b:4,3-b']dipyrrol-3(2H)-yl]carbonyl]-1H-indol-5-yl]-1H-indole-5-carboxamide(Compound 12)

Part A-Preparation of5-[[(2-carboxy-1H-indol-5-yl)amino]carbonyl]-1H-indole-2-carboxylic acid

Indole-2,5-dicarboxylic acid (98 mg, 0.42 mmoles) and5-amino-indole-2-carboxylic acid ethyl ester (86 mg, 0.42 mM) arestirred at room temperature under nitrogen in dry DMF (2 mL). EDC (90mg, 0.47 mM) is added and left to react for 25 hr. The reaction mixtureis diluted with water and centrifuged and the supernatant is decanted.The residual solid is dissolved in acetone and re-evaporated and then ischromatographed over silica gel (20 g), and eluted with 50% ethylacetate in hexane followed by 30% acetone in hexane. Fractions of 15 mlare collected, and analyzed by TLC.5-[[(2-carboxy-1H-indol-5-yl)amino]carbonyl]-1H-indole-2-carboxylic aciddiethyl ester is found in fractions 8-86 (137 mg, 78% yield).

Part B-Preparation of5-[[(2-carboxy-1H-indol-5-yl)amino]carbonyl]-1H-indole-2-carboxylic acid

5-[[(2-carboxy-1H-indol-5-yl)amino]carbonyl]-1H-indole-2-carboxylic aciddiethyl ester (137 mg, 0.33 mM) is dissolved with stirring at roomtemperature under nitrogen in pyridine (3 mL) and absolute ethanol (3mL). 1N NaOH (1 mL) is added and the reaction left to react for 29 hrduring which time a precipitate forms. 1N HCl (1 mL) is added and thereaction mixture concentrated on a rotary evaporator. Water is added tothe residue, forming a cloudy solution. The solution is freeze-dried.The residue is coated on celite (1 g) which is then added to the top ofa C-18 reversed phase silica gel column (10 g). The column is elutedwith 40% DMF water to 80% DMF water in 10% increments (100 mL each).Fractions of 5 mL are collected.5-[[(2-carboxy-1H-indol-5-yl)amino]carbonyl]-1H-indole-2-carboxylic acidis found in fractions 30-70 (40 mg).

Part C-Reaction of5-[[(2-carboxy-1H-indol-5-yl)amino]carbonyl]-1H-indole-2-carboxylic acidwith (Boc)CPI phenol chloride

(S)-1-(chloromethyl)-1,6-dihydro-5-hydroxy-8-methyl-benzo[1,2-b:4,3-b']dipyrrole-3(2H)-carboxylicacid 1,1-dimethyl ester [(Boc)CPI phenol chloride] (74 mg, 0.22 mM) isstirred at room temperature under nitrogen in dark for 45 min in ethylacetate (1 mL) and the ethyl acetate saturated with gaseous HCl (5 mL).The reaction mixture is evaporated under vacuum, and re-evaporated withCH₂ Cl₂ 2×, letting in nitrogen each time leaving(S)-1-(chloromethyl)-5-hydroxy-8-methyl-1,2,3,6-tetrahydro-benzo[1,2-b:4,3-b']dipyrrolehydrochloride (CPI phenol chloride hydrochloride). The CPI phenolchloride hydrochloride salt is dissolved with stirring under nitrogen indark in dry DMA (1 mL).5-[[(2-carboxy-1H-indol-5-yl)amino]carbonyl]-1H-indole-2-carboxylic acid(40 mg, 0.11 mM) and EDC (75 mg, 0.39 mM) are added to the reaction andleft to react for 30 min, when additional EDC (20 mg, 0.10 mM) is addedand left to react for 1 hr more. The solution is transferred to acentrifugal tube and washed in with DMF (1 mL), diluted with water toprecipitate product, and centrifuged. The supernatant is decanted. Thesolid is washed into RB flask with acetone and re-evaporated undervacuum.

The crude product is coated on silica gel (1 g) and placed on top of asilica gel column (9 g) made up in 20% DMF-80% toluene. The column iseluted with 20% DMF in toluene, followed by 30% DMF in toluene.Fractions of 2 ml are collected. Fractions 21-78 are collected andevaporated and the residue rechromatographed over a silica gel column (5g) eluted with 20% DMF in toluene. Fractions of 1 mL are collected.[S-(R*,R*)]-2-[[1-(chloromethyl)-1,6-dihydro-5-hydroxy-8'methyl-benzo[1,2-b:4,3-b']dipyrrol-3(2H)-yl]-carbonyl]-N-[2-[[1-(chloromethyl)-1,6-dihydro-5-hydroxy-8-methylbenzo[1,2-b:4,3-b']dipyrrol-3(2H)-yl]carbonyl]-1H-indol-5-yl]-1H-indole-5-carboxamideis found in fractions 20-45 (49 mg, 56% yield).

NMR(d₆ -DMSO,TMS): δ 2.37(s, 6H); 3.54-3.68(t, 2H); 3.87-3.97(d, 2H);3.98-4.10((t, 2H); 4.50-4.60(d, 2H); 4.61-4.77(q, 2H); 7.07(s, 2H);7.13(s, 1H); 7.29(s, 1H); 7.45-7.52(d, 1H); 7.55-7.73(m, 4H); 7.91(s,1H); 8.23(s, 1H); 8.45(s, 1H); 9.82(s, 1H); 9.84(s, 1H); 10.18(s, 1H);10.75(s, 1H); 10.76(s, 1H); 11.65(s, 1H); 12.00(s, 1H). UV(DMA+MeOH):λmax=328 nm (ε=36,830); 292nm sh (ε=43,800)

EXAMPLE 13 Preparation of[S-(R*,R*)]-5-[[1-(chloromethyl)-1,6-dihydro-5-hydroxy-8-methylbenzo[1,2-b:4,3-b']dipyrrol-3(2H)-yl]-carbonyl)-N-[2-[[1-(chloromethyl)-1,6-dihydro-5-hydroxy-8-methylbenzo[1,2-b:4,3-b']dipyrrol-3(2H)-yl]carbonyl)-1H-indol-5-yl]-1H-indole-2-carboxamide(Compound 13).

Part A-Preparation of5-[[(5-carboxy-1H-indol-2-yl)carbonyl]amino]-1H-indole-2-carboxylic aciddiethyl ester

Indole-2,5-dicarboxylic acid 5-ethyl ester (65 mg, 0.28 mM) is stirredat room temperature under nitrogen in dry DMF(1 mL).5-Amino-indole-2-carboxylic acid (57 mg, 0.28 mM) and EDC (60 mg, 31 mM)are added. After 24 hr the reaction is diluted with water and the solidscollected by filtration. The residual solid is dissolved in acetone andevaporated onto silica gel (2 g). This material is added to the top of asilica gel column (18 g) and eluted with 10% DMF in toluene.5-[[(5-carboxy-1H-indol-2-yl)carbonyl]amino]-1H-indole-2-carboxylic aciddiethyl ester (75 mg, 64% yield) is found in fractions.

Part B-Preparation of5-[[(5-carboxy-1H-indol-2-yl)carbonyl]amino]-1H-indole-2-carboxylic acid

5-[[(5-carboxy-1H-indol-2-yl)carbonyl]amino]-12H-indole-2-carboxylicacid diethyl ester (75 mg, 0.18 mM) is stirred at room temperature undernitrogen in pyridine (2 mL), absolute ethanol (2 mL) and 1N NaOH (600μL) for 120 hr at 25° C. Additional 1N NaOH (1 mL) is then added and thereaction heated for 20 hr at 50° C. 1N HCl (1.6 mL) is then added andthe reaction evaporated under vacuum. The residue is coated on Celite (1g) and placed in an C-18 silica gel column (10 g). The column is elutedwith 508 DMF in water to 80% DMF in water (100 ml each in 108increments). Fractions of 5 ml are collected and analyzed by TLC.5-[[(5-carboxy-1H-indol-2-yl)carbonyl]amino]-1H-indole-2-carboxylic acidis found in fractions 5-77.

Part C-Reaction of5-[[(5-carboxy-1H-indol-2-yl)carbonyl]amino]-1H-indole-2-carboxylic acidwith (Boc)CPI phenol chloride

(S)-1-(chloromethyl)-1,6-dihydro-5-hydroxy-8-methyl-benzo[1,2-b:4,3-b']dipyrrole-3(2H)-carboxylicacid 1,1-dimethyl ester [(Boc)CPI phenol chloride] (121 mg, 0.36 mM) isstirred at room temperature under nitrogen in dark for 30 min in ethylacetate (2 mL) and ethyl acetate saturated with gaseous HCl (8 mL). TLCafter 30 min shows the reaction to be complete. The reaction isevaporated under vacuum and re-evaporated with CH₂ Cl₂ 2×, letting innitrogen each time giving(S)-1-(chloromethyl)-5-hydroxy-8-methyl-1,2,3,6-tetrahydro-benzo[1,2-b:4,3-b']dipyrrolehydrochloride (CPI phenol chloride hydrochloride). The CPI phenolchloride hydrochloride salt is dissolved with stirring under nitrogen inthe dark in dry DMA (1.5 mL).5-[[(5-carboxy-1H-indol-2-yl)carbonyl]amino]-1H-indole-2-carboxylic acid(65 mg0 0.18 mM) and EDC (110 mg, 0.57 mM) are added to the resultantsolution. After 30 min reation is treated with additional EDC (45 mg,0.23 mM) and left to react 1.5 hr more. The reaction solution istransferred to a centrifuge tube and washed in with DMF (1 mL), dilutedwith water to precipitate product, and centrifuged. The supernatant isdecanted. The solid is washed into RB flask with acetone andre-evaporated under vacuum. The crude product is coated on silica gel (1g) and placed on top of a silica gel column (10 g) made up in 20%DMF-808 toluene. The column is eluted with 20% DMF in toluene, followedby 358 DMF in touene. Fractions of 2 ml are collected.[S-(R*,R*)]-5-[[1-(chloromethyl)-1,6-dihydro-5-hydroxy-8-methylbenzo[1,2-b:4,3-b']dipyrrol-3(2H)-yl]-carbonyl)-N-[2-[[1-(chloromethyl)-1,6-dihydro-5-hydroxy-8-methyl-benzo[1,2-b:4,3-b']dipyrrol-3(2H)-yl]carbonyl)-1H-indol-5-yl]-1H-indole-2-carboxamide(43 mg) is isolated from fractions 25-47.

NMR(d₆ -DMSO,TMS): δ 2.33(s, 3H); 2.37(s, 3H); 3.50-3.70(t, 2H);3.78-3.97(m, 3H); 3.97-4.14(t, 2H); 4.24-4.43(bs, 1H); 4.48-4.62(d, 1H);4.62-4.75(t, 1H); 7.04(s, 1H); 7.06(s, 1H); 7.16(s, 1H); 7.40-7.72(m,7H); 8.00(s, 1H); 8.25(s, 1H); 9.64-9.80(bs, 1H); 9.81(s, 1H); 10.30(s,1H); 10.68(s, 1H); 10.75(s, 1H); 11.70(s, 1H); 12.03(s, 1H).UV(DMA+MeOH): λmax=316 nm sh (ε 36,830); 295 nm (ε=40,030)

EXAMPLE 14 Preparation of[S-(R*,R*)]-carbonylbis[imino-1H-indole-5,2-diylcarbonyl[1-(chloromethyl)-1,6-dihydro-8-methylbenzo[1,2-b:4,3-b']dipyrrole-3,5(2H)-diyl]]-ester,2,2-dimethylpropanoic acid (Compound 14)

[7bR-[2(7'bR*,8'aS*),7bR*,8aS*]]-2,2'-[carbonylbis(imino-1H-indole-5,2-dicarbonyl)]bis[1,2,8,8a-tetrahydro-7-methyl-cyclopropa(c)pyrrolo[3,2-e]indol-4(5H)-one(5.4 mg, 7.3 μM) is dissolve in pyridine (200 μL) and the the solutiontreated with 2,2-dimethylpropionic acid chloride (24 μL, 192 μM) at 0 C.After 10 minutes the reaction is warmed to 25 C and is stirred 20 min atthat temperature. The reaction is then quenched with water (0.1 mL) andconcentrated to dryness. The residue is dissolved in DMF and evaporatedunder vacuum onto silica gel (0.1 g). This material is added to the topof a silica gel column (1 g) and eluted with 10% to 30% DMF in toluene.Fractions of 0.2 mL are collected. Fractions 17-50 are evapoarated todryness and the residue washed with acetone. The residue from theacetone wash is then chromatographed over a silica gel column (0.3 g)eluted with 10%-12% DMF in toluene. Fractions of 0.2 mL are collected.[S-(R*,R*)]-carbonylbis[imino-1H-indole-5,2-diylcarbonyl[1-(chloromethyl)-1,6-dihydro-8-methylbenzo[1,2-b:4,3-b']dipyrrole-3,5(2H)-diyl]]ester,2,2-dimethylpropanoic acid (1.1 mg) is obtained on evaporation of fr9-23.

NMR(d₆ -DMSO,TMS): δ 1.39(s, 18H); 2.41 (s, 6H); 3.73 (m, 2H); 4.00 (m,2H); 4.21 (m, 2H); 4.60 (m, 2H); 4.73(m, 2H); 7.09 (s, 2H); 7.25 (m,4H); 7.42 (m, 2H); 7.80 (bs, 2H); 7.88 (s, 2H); 8.55 (bs, 2H); 10.87(bs, 2H); 11.54 (bs, 2H). UV(DMA+MeOH): λmax=324 nm (ε=40,000); 294 nm(ε=40,000).

EXAMPLE 15 Preparation of[S-(R*,R*)]-carbonylbis[imino-1H-indole-5,2-diylcarbonyl[1-(chloromethyl)-1,6-dihydro-

8-methylbenzo[1,2-b:4,3-b']dipyrrole-3,5(2H)-diyl]]-ester, decanoic acid(Compound 15).

[S-(R*,R*)]-6,6'-[carbonylbis(imino-1H-indole-5,2-dicarbonyl)]-bis[8-chloromethyl)-3,6,7,8-tetrahydro-1-methyl-benzo[1,2-b:4,3-b']dipyrrol-4-ol(10 mg, 0.012 mM) is dissolved in pyridine (0.2 mL) and DMF (0.01 mL).The solution is treated with decanoic anydride (75 mg, 0.23 mM) and thereaction stirred in the dark for 24 hr. The reaction is then quenchedwith water (0.05 mL) and evaporated to dryness. The residue is washedwith water and with toluene each the insolubles are collected bycentrifugation. The resultant residue is chromatographed over a silicagel column (1.2 g) eluted with 5%, 6%, 7%, and 10% DMF in toluene.Fractions of 0.3 mL are collected.[-S(R*,R*)]-carbonylbis[imino-1H-indole-5,2-diylcarbonyl[1-(chloro-methyl)-1,6-dihydro-8-methylbenzo[1,2-b:4,3-b']dipyrrole-3,5(2H)-diyl]]ester,decanoic acid (1.3 mg) is obtained on evaporation of fractions 18-60.

NMR(d₆ -DMSO,TMS): δ 0.86 (t); 1.24 (m); 1.45 (m); 2.18 (t); 2.40(s);3.70 (m); 3.98(m); 4.15 (m); 4.59 (m); 4.71 (m); 7.08 (s); 7.27 (m);7.42 (m); 7.80 (m); 7.88 (s); 8.59 (bs); 11.05 (bs); 11.53 (bs).UV(DMA+MeOH): λmax=324 nm (ε=40,000); 294 nm (ε=40,000).

EXAMPLE 16 Preparation of[S-(R*,R*)]-6,6'-[carbonylbis[(7,8-dihydrobenzo[1,2-b:4,3-b']dipyrrole-6,2(3H)-diyl)carbonyl]]bis[8-(chloromethyl)-3,6,7,8-tetrahydro-1-methyl-benzo[1,2-b:4,3b']dipyrrol-4-ol(Compound 16)

Part A-Preparation of6,6'-carbonylbis[3,6,7,8-tetrahydrobenzo[1,2-b:4,3-b']dipyrrole-2-carboxylicacid ethyl ester is described in M. A. Warpehoski, V. S. Bradford,Tetrahedron Lett, 1986, 27, 2735.

NMR: δ 1.35(t, 6H) 3.285(t, 4H); 4.123(t, 4H); 4.34(q, 4H); 7.06(s, 2H);7.22(d, 2H); 7.32(d, 2H); 11.845(s, 2H). MS(E1): m/z 586(M+), 257, 230,184, 156. TLC: R_(f) =0.14 in (25-75) Acetone-hexane.

Part B-Preparation of6,6'-carbonylbis[3,6,7,8-tetrahydrobenzo[1,2-b:4,3-b']dipyrrole-2-carboxylicacid diethyl ester.

6,6'-carbonylbis[3,6,7,8-tetrahydro-benzo[1,2-b:4,3-b']dipyrrole-2-carboxylicacid ethyl ester (30 mg, 0.13 mM) is dissolve in dry THF (0.5 mL) andthe solution treated with 4-dimethylaminopyridine (1 mg). This solutionis cooled to -98° C. in a liquid nitrogen-frozen methanol bath undernitrogen. Diisopropylethylamine is added (25 μl, 0.14 mM) followed by1.93M phosgene in toluene (35 μl, 0.067 mM). The reaction is stirred atabout -98° C. for 3 hr. It is then stored in the -65° C. freezerovernight. The reaction is allowed to stir at room temperature for 90min. It is diluted with water and ethyl acetate. The ethyl acetate layeris washed with brine, dried and evaporated. The crude residue (47.9 mg)is adsorbed onto silica gel (0.5 g) from distilled THF. The resultantsolid is added to the top of a silica column (5 g) and eluted with5%-15% acetone in toluene. Fractions are of 3 mL are collected.6,6'-carbonylbis[3,6,7,8-tetrahydrobenzo[1,2-b:4,3-b']dipyrrole-2-carboxylicacid diethyl ester (5.6 mg) is found in fraction 5-9.

PartC-6,6'-carbonylbis[3,6,7,8-tetrahydrobenzo[1,2-b:4,3-b']dipyrrole-2-carboxylicacid diethyl ester (7 mg, 0.014 mM) is dissolved in pyridine and thesolution treated with 1N NaOH (0.05 mL). The reaction is then heated to50 C for 6 hr. The reaction is acidified with 1N HCl (0.2 mL) and6,6'-carbonylbis[3,6,7,8-tetrahydrobenzo[1,2-b:4,3-b']dipyrrole-2-carboxylicacid diethyl ester precipitated as a solid is collected bycentrifugation and dried under vacuum.

Part D-Preparation of[S-(R*,R*)]-6,6'-[carbonylbis[(7,8-dihydrobenzo[1,2-b:4,3-b']dipyrrole-6,2(3H)-diyl)carbonyl]]bis[8-(chloromethyl)-3,6,7,8-tetrahydro-1-methyl-benzo[1,2-b:4,3b']dipyrrol-4-ol(Compound 16).

(S)-1-(chloromethyl)-1,6-dihydro-5-hydroxy-8-methyl-benzo[1,2b:4,3-b']dipyrrole-3(2H)-carboxylic acid 1,1-dimethyl ester [(Boc)CPI phenol chloride] (11 mg,0.033 mM) is dissolved in ethyl acetate (0.2) and HCl-saturated ethylacetate (0.7 mL). The mixture is stirred at room temperature undernitrogen for 1 hr. It is evaporated under a nitrogen stream. CH₂ C₁₂ isadded, and the residual(S)-1-(chloromethyl)-5-hydroxy-8-methyl-1,2,3,6-tetrahydrobenzo[1,2-b:4,3-b']dipyrrolehydrochloride (CPI phenol chloride hydrochloride salt) is re-evaporatedtwice under a nitrogen stream.6,6'-carbonylbis[3,6,7,8-tetrahydrobenzo[1,2-b:4,3-b']dipyrrole-2-carboxylicacid diethyl ester (6 mg, 0.014 mM) is dissolved in DMA (0.3 mL) andadded to the CPI phenol chloride hydrochloride salt. Then the mixture istreated with EDC (6.4 mg, 0.034 mM). The reaction is stirred undernitrogen at room temperature 50 min. Additional EDC (6.4 mg, 0.034 mM)is added, and stirring is continued 65 min. The product is precipitatedout by the gradual addition of water. The solids (28 mg) are collectedby centrifugation and dried under vacuum. This crude product is adsorbedfrom DMF onto silica gel (0.3 g). It is put on a silica gel column (3g), eluting with 10, 12, 14, 16 and 20% DMF-toluene. Fractions of 1 mLare collected. [S-(R*,R*)]-6,6'-[carbonylbis[(7,8-dihydrobenzo[1,2-b:4,3-b']dipyrrole-6,2(3H)-diyl)carbonyl]]bis[8-(chloromethyl)-3,6,7,8-tetrahydro-1-methyl-benzo[1,2-b:4,3b']dipyrrol-4-ol(6 mg) is obtained on concentration of fractions 40-62.

NMR: δ 2.36(s, 6H); 3.3(t, 4H); 3.61(m, 2H); 3.92(m, 2H); 4.03(m, 2H);4.16(t, 4H); 4.55(m, 2H); 4.69(m, 2H); 7.01(s, 2H); 7.05(s, 2H); 7.27(m,4H); 7.66(m, 2H); 9.79(s, 2H); 10.74(bs, 2H); 11.62(bs, 2H). UV:λmax=353 nm (ε=28000); 286 nm(ε=45000). TLC: R_(f) =0.29 in (20-80)DMF-toluene.

The starting compounds are known or can be readily prepared by knownmethods. See M. A. Warpehoski, Tet. Lett., 27, 4103 (1986); W. W.Wierenga, J. Am. Chem. Soc., 103, No. 18, 1981; D. G. Martin, J.Antibiotics 1985, 38, 746; and M. A. Warpehoski, I. Gebhart, R. C.Kelly, W. C. Krueger, L. H. Li, J. P. McGovren, M. D. Prairie, N.Wicnienski and W. Wierenga, J. Med. Chem., 1988, 31, pp. 590-603.

The preparation of CPI(Boc) HCl is described in R. C. Kelly, I. Gebhard,N. Wicnienski, P. A. Aristoff, P. D. Johnson, D. G. Martin, J. Am. Chem.Soc. 1987, 109 6837.

The spirocyclopropylcyclohexadienyl compounds of Formula A and1-(halomethyl)-1,6-hydro-5-hydroxy-8-methyl-benzo[1,2-b:4,3-b']dipyrrole-3(2H)-yl5-ester or urethanes (Formula B) can also be prepared by the proceduresand methods disclosed in U.S. patent application Ser. No. 894,314, filedAug. 7, 1986 (now abandoned), and PCT/87/03227 patent application filedDec. 11, 1987. Both are incorporated herein by reference. See also EPApplication 0 154 445 (published 9 Nov. 1985).

All the compounds of the subject invention have UV absorption in therange of 200 nm to 380 nm. Thus, novel compounds of the subjectinvention (Formula I) are useful as UV adsorbents in technical andindustrial areas, as follows:

(a) textile materials, for example, wool, silk, cotton, hemp, flax,linen and the like;

(b) natural or synthetic resins.

Depending on the nature of the material to be treated, the requirementswith regard to the degree of activity and durability, and other factors,the proportion of the light screening agent to be incorporated in thematerial may vary within fairly wide limits, for example, from about0.01% to about 10%, and, advantageously, 0.1% to 2% of the weight of thematerial which is to be directly protected against the action of UVrays.

The compounds of Formula I are particularly useful as antitumor agents.Examples of compounds of Formula I demonstrate antitumor activity inP388 leukemic mice, and also show significant activity in the L1210leukemia and B16 melanoma murine test systems. These murine test systemsare predictive for clinically useful human antitumor agents (see, forexample, A. Geldin et al, European J. Cancer, Vol. 17, pp 129-142, 1981;J. M Vendetti, Cancer Treatment Reports, Vol. 67, pp. 767-772, 1983; andJ. M. Vendetti et al, Advances in Pharmacology and Chemotherapy, Vol.20, pp. 1-20, 1984), and, therefore, the compounds of the subjectinvention (Formula I) will be useful in the control and treatment ofsusceptible neoplastic (cancer) diseases, including susceptibleleukemics, in humans when given, for example, intravenously in doses of0.001 μg/kg to about 10 mg/kg of body weight per day, the exact dosedepending on the age, weight, and condition of the patient, and on thefrequency of administration.

The compounds of Formula I are effective when administered intravenously(IV) in fluid solutions by bolus injection or by infusion. The preferreddoses are 0.01 microgram/kg to 1000 microgram/kg by bolus injection and0.0002 to 20 microgram/kg/min by infusion. The exact dose will varydepending on the particular compound as well as the age, weight, routeof administration, and physical condition of the patient, and on thefrequency of administration.

Illustrative in vivo and in vitro L1210 testing data on the compounds ofFormula I are presented in Tables 1 and 2. Table 3 presents datacomparing[S-(R*,R*)]-6,6'-[carbonylbis(imino-1H-indole-5,2-dicarbonyl)]bis[8-chloromethyl)-3,6,7,8-tetrahydro-1-methyl-benzo[1,2-b:4,3-b']dipyrrol-4-ol(Compound 1) with(7bR)-N-[2-[(4,5,8,8a-tetrahydro-7-methyl-4-oxocyclopropa-[c]pyrrolo[3,2-e]-indol-2(1H)Oyl)carbonyl]-1H-indol-5-yl]-2-benzofurancarboxamide(U-73,975) in the other murine systems where the compounds have been runjointly. Compound 1 is as least as active as the compound U-73,975 inevery system and in some such as the subcutaneous L1210 assay it showssuperior activity over several dose ranges. Further, Compound 1, likeU-73,975, has been found not to cause delayed death.

In vivo L1210 biological data shows[S-(R*,R*)]-6,6'-[carbonyl-bis(imino-1H-indole-5,2-dicarbonyl)]bis[8-chloromethyl)-3,6,7,8-tetrahydro-1-methyl-benzo[1,2-b:4,3-b']dipyrrol-4-ol(Compound 1) to be the most active analog being highly curative at 15μg/kg. The results set forth in Tables 1 and 4 were obtained usingstandard well known procedures (In Vivo Cancer, Models, NIH PublicationNo. 84-2635, 1984).

T/C refers to median life span of treated mice divided by median lifespan of control mice times 100.

The compounds of formula I are useful as antibacterial agents. Thesecompounds are useful to control the proliferation of susceptiblemicrobes in various environments using standard microbiologicaltechniques. Such environments include dental utensils contaminated withS. aureus, and the like, and laboratory benches in a microbiologicallaboratory which can be cleansed with a formulation containing about1-10% (w/v) of a compound of formula I. ##STR1##

SUPPLEMENTAL FORMULA CHART

-het- is selected from the group consisting of: ##STR2##

where X₁₂ and and X₁₃ are the same or different and are H, halogen, C₁-C₅ alkyl, or NO₂ ; ##STR3##

where X₁₂ and X₁₃ are as defined above and R₁ is C₁ -C₅ alkyl; ##STR4##

where X₁₂ and X₁₃ are as defined above; ##STR5##

where X₁₂ and R₁ are as defined above; ##STR6##

where X₁₂ is as defined above and X₈ is --O--, --S--, --NH--; ##STR7##

wherein X₈ is as defined above and X₉ is --CH═ or --N═; ##STR8##

where X₉, X₁₂ and X₁₃ are as defined above; ##STR9##

where X₂ and X₁₃ are as defined above, and when X₉ and X₁₀ are the sameor different, and are selected from --CH═ or --N═; ##STR10##

wherein X₉, X₁₀, X₁₂, X₁₃ are as defined above; ##STR11##

where X₉, X₁₀, X₁₂ and X₁₃ are as defined above; ##STR12##

wherein X₈, X₉, X₁₂ and X₁₃ are as defined above;

CHART C ##STR13##

where X₁ is H, CH₃, OH, OCH₃, NO₂, NH₂, (NHNHAc) NHNHC(O)CH₃, (NHBz)NHC(O)C₆ H₅, or halogen; ##STR14##

where X₃ is H, OH or OCH₃ ; ##STR15##

where R₈ is H, CH₃ or C₂ H₅ ; ##STR16##

wherein R' is H or CH₃ S--; ##STR17##

where X₈ is --O--, --S--, NH; X₉ is --CH═ or --N═; ##STR18##

where X₉ and X₈ have the meanings defined above; and Y₁ is H, halo, C₁-C₄ -alkyl, C₁ -C₃ -alkoxy, C₂ -C₆ -dialkyl amino, nitro, amino-carbonylalkyl(C₁ -C₁₀), hydroxy, amino (--NH₂), --NHCONH₂, --NHAc (NHCOCH₃) or--NHBz (NHC(O)--C₆ H₅); ##STR19##

where X₈ and Y₁ have the meanings defined above; ##STR20##

where X₁₀ is --CH═ or --N═; ##STR21##

where X₁₀ has the meanings defined above; and ##STR22##

where R₈ has the meanings defined above, ##STR23##

wherein Y₁ and X₈ have the meanings defined above; ##STR24##

wherein Y₁, X₈ and X₉ have the meanings defined above; ##STR25##

wherein X₁₀ is --CH═ or --N═, and Y₁ have the meanings defined above;##STR26##

wherein and Y₁ has have the meanings defined above.

                  TABLE 1                                                         ______________________________________                                        L-1210 Leukemia in vivo                                                       IV drug vs IP tumor dosed on day 1                                            L1210 In vivo                                                                 Compound # Dose (μg/kg) Best T/C.sup.b                                     ______________________________________                                        14         20              156                                                 9         50              165                                                10          6              213 (1 cure)                                       11         13              188                                                12          2              181                                                13         13              188                                                U-73,975   100             213                                                 1         15              4/6 cures                                           3          6              106                                                 8         50              194                                                 2         10              169                                                 6          5              178 (1 cure)                                        7          1              156                                                 5         600             156                                                 4         50              156                                                15         13              243 (1 cure)                                       16         12              243 (1/6 cure)                                     ______________________________________                                         .sup.a Male BDF.sub.1 mice                                                    .sup.b T/C = Treated/Control × 100, where Treated is the median         survival time of the treated group and control is the median survival tim     of the untreated control group. Animals surviving 30 days are considered      cured.                                                                   

                  TABLE 2                                                         ______________________________________                                        In vitro Biological Data                                                      Compound #            ID.sub.50.sup.b                                         ______________________________________                                        1                     0.000001                                                3                     0.000009                                                8                     0.000006                                                2                     0.000001                                                6                     0.000012                                                7                     0.000004                                                5                     0.00001                                                 4                     0.000004                                                15                    0.000001                                                16                    0.000002                                                ______________________________________                                         .sup.a Drugs are tested against L1210 leukemia cells.                         .sup.b The concentration of drug in μg/ml which inhibits cell growth       50% is reported.                                                         

                  TABLE 3                                                         ______________________________________                                                       U-73-975 Cpd 1                                                 ______________________________________                                        L-1210 i.p..sup.a                                                                        OD.sup.b  100        10-15                                         i.v. dose  μg/kg                                                           on day 1   best T/C  213        (4/6)                                                    (cures).sup.c                                                      L-1210 i.p.sup.a                                                                         OD         20         4                                            i.p. dose  μg/kg                                                           on days 1, 5, 9                                                                          T/C       (4/6)      (3/6)                                                    (cures)                                                            L-1210 sc  OD        100        10-25                                         i.p. dose  μg/kg                                                           on day 1   best T/C  (4/6)      (6/6)                                         B-16 i.p.  OD        100        15                                            i.v. dose  μg/kg                                                           on day 1   T/C       160        167                                           Lewis Lung i.v.                                                                          OD         25         4                                            i.v. dose μg/kg                                                            on days 1, 5, 9                                                                          T/C       147        168                                           NO TUMOR             No Delayed No Delayed                                                         Death      Death                                         ______________________________________                                         .sup.a i.p. = intraperitoneally and i.v. = intravenously                      .sup.b Od = optimum dose                                                      .sup.c In these tests, animals which survive 30 days are considered cured     ##STR27##

What is claimed is:
 1. A compound of formula I

    CPI.sub.1 -R.sub.5 T-R'.sub.5 -CPI.sub.2

wherein CPI₁ and CPI₂, being the same or different, are selected fromFormula A or B ##STR28## wherein W is selected from C₁ -C₅ alkyl, phenylor hydrogen; wherein X is selected from azido, a halogen atom, cyanate,thiocyanate, isocyanate, thioisocyanate, phosphate diester (--PO(OR)₂),phosphonyl (--O--PO₂ R), thiophosphonyl (--O--PSOR), sulfinyl (--O--SOR)or sulfonyl (--O--SO₂ R); wherein Y is selected from hydrogen, --C(O)R,--C(S)R, --C(O)OR₁, --S(O)₂ R₁, --C(O)NR₂ R₃, --C(S)NR₂ R₃, or--C(O)NHSO₂ R₄ ; wherein Z is selected from the group consisting of C₁-C₅ alkyl, phenyl or hydrogen; wherein R is selected from the groupconsisting of C₁ -C₂₀ alkyl; C₂ -C₆ alkenyl; C₂ -C₆ alkynyl; phenyloptionally substituted with one, 2 or 3 C₁ -C₄ alkyl, C₁ -C₃ alkoxy,halo, C₁ -C₃ alkylthio, trifluoromethyl, C₂ -C₆ dialkylamino, or nitro;naphthyl optionally substituted with one or 2 C₁ -C₄ alkyl, C₁ -C₃alkoxy, halo, trifluromethyl, C₂ -C₆ dialkylamino, C₁ -C₃ alkylthio ornitro; wherein R₁ is selected from C₁ -C₂₀ alkyl or phenyl optionallysubstituted with one, 2 or 3 C₁ -C₄ alkyl, C₁ -C₃ alkoxy, halo, C₁ -C₃alkylthio, trifluoromethyl, C₂ -C₆ dialkylamino, or nitro; wherein R₂and R₃, being the same or different, are selected from hydrogen, C₁ -C₂₀alkyl, or phenyl optionally substituted with one, 2 or 3 C₁ -C₄ alkyl,C₁ -C₃ alkoxy, halo, C₁ -C₃ alkylthio, trifluoromethyl, C₂ -C₆dialkylamino, or nitro; with the proviso that both R₂ and R₃ can not bephenyl or substituted phenyl; wherein R₄ is selected from C₁ -C₁₀ alkyl;phenyl optionally substituted with one, 2 or 3 C₁ -C₄ alkyl, C₁ -C₃alkoxy, halo, C₁ -C₃ alkylthio, trifluoromethyl, C₂ -C₆ dialkylamino, ornitro; naphthyl optionally substituted with one or 2 C₁ -C₄ alkyl, C₁-C₃ alkoxy, halo, trifluromethyl, C₂ -C₆ dialkylamino, C₁ -C₃ alkylthioor nitro; wherein R₅ and R'₅, being the same or different, are selectedfrom a direct bond or a carbonyl acyl group selected from the groupconsisting of: ##STR29## where X₁ is H, CH₃, OH, OCH₃, NO₂, NH₂,(NHNHAc) NHNHC(O)CH₃, (NHBz) NHC(O)C₆ H₅, or halogen; ##STR30## where X₃is H, OH or OCH₃ ; ##STR31## where R₈ is H, CH₃ or C₂ H₅ ; ##STR32##wherein R' is H or CH₃ S--; ##STR33## where X₈ is --O--, --S--, NH; X₉is --CH═ or --N═; ##STR34## where X₉ and X₈ have the meanings definedabove: and Y₁ is H, halo, C₁ -C₄ -alkyl, C₁ -C₃ -alkoxy, C₂ -C₆-dialkylamino, nitro, amino-carbon ylalkyl(C₁ -C₁₀), hydroxy, amino(--NH₂), --NHCONH₂, --NHAc (NHCOCH₃) or --NHBz (NHC(O)--C₆ H₅);##STR35## where X₈ and Y₁ have the meanings defined above; ##STR36##where X₁₀ is --CH═ or --N═; ##STR37## where X₁₀ has the meanings definedabove; and ##STR38## where R₈ has the meanings defined above; ##STR39##wherein Y₁ and X₈ have the meanings defined above; ##STR40## wherein Y₁,X₈ and X₉ have the meanings defined above; ##STR41## wherein X₁₀ is--CH═ or --N═, and Y₁ have the meanings defined above; ##STR42## whereinY₁ has the meanings defined above; wherein T is a tether linkageselected from the group consisting of:a) amino carbonyl (--NHC(O)--); b)carbonylamino (--C(O)NH--); c) carbonyloxy (--C(O)O--); d) oxycarbonyl(--OC(O)--); e) amino-tether-amino of the formula -NR₁₃ -T'-NR₁₄ - whereR₁₃ and R₁₄, being the same or different, are hydrogen or C₁ -C₈ alkyl;or when taken together are --(CH₂)_(n) -- where n is 2 or 3: where T' isselected from the group consisting of carbonyl (--C(O)--), dicarbonyl(--C(O)C(O)--); (--C(O)(CH₂)_(n) C(O)--), where n is 1 to 5,(--C(O)PhC(O)--), where Ph is 1,3- or 1,4- phenylene, or a group of theformula --C(O)-het-C(O)--; f) --C(O)-het-C(O)--, when R₅ and R'₅ areboth a direct bond, wherein -het- is a fused mono-, di-, or tricyclicheteroaryl of 5 to 12 members, containing one, two, or three heteroatomsselected from the group consisting of oxygen, nitrogen or sulfur,optionally substituted with one or 2 C₁ -C₄ alkyl, C₁ -C₃ alkoxy, halo,C₁ -C₃ alkylthio, trifluoromethyl, C₂ -C₆ dialkylamino, or nitro.
 2. Acompound according to claim 1 wherein W is methyl.
 3. A compoundaccording to claim 1 wherein X is halogen.
 4. A compound according toclaim 1 wherein Y is hydrogen or selected from the group consisting of--COR, wherein R is selected from C₁ -C₁₀ alkyl.
 5. A compound accordingto claim 1 wherein Z is hydrogen.
 6. A compound according to claim 1wherein T is selected from the the group consisting of an amide(aminocarbonyl); carbonylamino (--C(O)NH--); or an amino-tether-amino ofthe formula -NR₁₃ -T'-NR₁₄ - where R₁₃ and R₁₄ are hydrogen and where T'is carbonyl (--C(O)--) or is selected from a group of the formula--C(O)-het-C(O)--.
 7. A compound according to claim 1 wherein T isselected from the group consisting of an amino-tether-amino of theformula -NR₁₃ -T'-NR₁₄ - where R₁₃ and R₁₄ are hydrogen and where T' iscarbonyl (--C(O)--) or is selected from a group of the formula--C(O)-het-C(O)-- wherein -het- is a heteroaryl selected frompyrrol-2,5-diyl, fur-2,5-diyl, indol-2,5-diyl, benzofuran-2,5-diyl or3,6-dihydrobenzo[1,2-b:4,3-b']dipyrrol-2,7-diyl.
 8. A compound accordingto claim 1 wherein R₅ and R'₅ are selected from the group consisting of2-carbonylindole-5-yl, 2-carbonyl-6-hydroxy-7-methoxyindol-5-yl,2-carbonyl-1,2,3,6-tetrahydrobenzo[1,2-b:4,3-b']dipyrrol-7-yl,2-carbonyl-4-hydroxy-5-methoxy-1,2,3,6-tetrahydrobenzo[1,2-b:4,3-b']dipyrrol-7-yl.9. A compound according to claim 1 wherein CPI₁ and CPI₂, being the sameor different, are preferably1-(chloromethyl)-1,6-dihydro-8-methyl-5-hydroxy-benzo[1,2:4,3-6']dipyrrole-3(2H)-yland4,5,8,8a-tetrahydro-7-methyl-4-oxocyclopropa[c]pyrrolo(3,2-e)indol-2(1H)-yl.10. A compound according to claim 1 selected from the group consistingof:[S-(R*,R*)]-6,6'-[carbonylbis(imino-1H-indole-5,2-dicarbonyl)]-bis[8-chloromethyl)-3,6,7,8-tetrahydro-1-methyl-benzo[1,2-b:4,3-b']dipyrrol-4-ol(Compound 1); [7bR-[2(7'bR*,8'aS*),7bR*,8aS*]]-2,2'[carbonylbis(imino-1H-indole-5,2-dicarbonyl)]bis[1,2,8,8a-tetrahydro-7-methyl-cyclopropa(c)pyrrolo[3,2-e]indol-4(5H)-one(Compound 2);(R*,S*)-N,N'-bis[2-[[1-(chloromethyl)-1,6-dihydro-5-hydroxy-8-methylbenzo[1,2-b:4,3-b']dipyrrol-3(2H)-yl]carbonyl]-1H-indol-5-yl]-ethanediamide(Compound 3); [S-(R*,R*)]-6,6'-(1H-pyrrole-2,5-diyldicarbonyl)bis[8-(chloromethyl)-3,6,7,8-tertrahydro-1-methyl-benzo[1,2-b:4,3-b'-]dipyrrol-4-ol(Compound 4):[S-(R*,R*)]-6,6'-(2,5-furandiyldicarbonyl)bis[8-(chloromethyl)-3,6,7,8-tetrahydro-1-methyl-benzo[1,2-b:4,3'b]dipyrrol-4-ol(Compound 5);[S-(R*,R*]-N,N'-bis[2-[[1-(chloromethyl)-1,6-dihydro-5-hydroxy-8-methylbenzo[1,2-b:4,3-b']dipyrrol-3(2H)-yl]carbonyl]-1H-indol-5-yl]-2,5-furandicarboxamide(Compound 6);[S-(R*,R*)]-N,N'-bis[2[[1-(chloromethyl)-1,6-dihydro-5-hydroxy-8-methylbenzo[1,2-b:4,3-b']dipyrrol-3(2H)-yl]carbonyl]-1H-indol-5-yl]-1H-pyrrole-2,5-dicarboxamide(Compound 7);[R-(R*,S*)]-N,N'-bis[2-[[1-(chloromethyl)-1,6-dihydro-5-hydroxy-8-methylbenzo[1,2-b:4,3-b']dipyrrol-3(2H)-yl]carbonyl]-1H-indol-5-yl]-propanediamide (Compound 8);[S-(R*,R*)]-6,6'-[carbonylbis(imino-1H-indole-5,2-diylcarbonyl)]bis[8-(chloromethyl)-3,6,7,8-tetrahydro-1-methyl-benzo[1,2b:4,3-b']dipyrrol-4-oldiacetate (Compound 9);[S-(R*,R*)]-6,6'-(1H-indole-2,5-diyldicarbonyl)bis[8-(chloromethyl)-3,6,7,8-tetrahydro-1-methyl-benzo[1,2-b:4,3-b']dipyrrol-4-ol(Compound 10);[S-(R*,R*)]-N,N'-bis[2-[[1-(chloromethyl)-1,6-dihydro-5-hydroxy-8-methylbenzo[1,2-b:4,3-b']dipyrrol-3(2H)-yl]carbonyl]-1H-indol-5-yl]-1H-indole-2,5-dicarboxamide(Compound 11);[S-(R*,R*)]-2-[[1-(chloromethyl)-1,6-dihydro-5-hydroxy-8-methyl-benzo[1,2-b:4,3-b']dipyrrol-3(2H)-yl]carbonyl]-N-[3-[[1-(chloromethyl)-1,6-dihydro-5-hydroxy-8-methylbenzo[1,2-b:4,3-b']dipyrrol-3(2H)-yl]carbonyl]-1H-indol-6-yl]-1H-indole-5-carboxamide (Compound 12);[S-(R*,R*)]-5-[[1-(chloromethyl)-1,6-dihydro-5-hydroxy-8-methylbenzo[1,2-b:4,3-b']dipyrrol-3(2H)-yl]-carbonyl)-N-[2-[[1-(chloromethyl)-1,6-dihydro-5-hydroxy-8-methylbenzo[1,2-b:4,3-b']dipyrrol-3(2H)-yl]carbonyl)-1H-indol-5-yl]-1H-indole-2-carboxamide(Compound 13);[S-(R*,R*)]-carbonylbis[imino-1H-indole-5,2-diylcarbonyl[1-(chloromethyl)-1,6-dihydro-8-methylbenzo[1,2-b:4,3-b']dipyrrole-3,5(2H)-diyl]]ester,2,2-dimethylpropanoic acid (Compound 14);[S-(R*,R*)]-carbonylbis[imino-1H-indole-5,2-diylcarbonyl[1-(chloromethyl)-1,6-dihydro-8-methylbenzo[1,2-b:4,3-b']dipyrrole-3,5(2H)-diyl]]ester,decanoic acid (Compound 15);[S-(R*,R*)]-6,6'-[carbonylbis[(7,8-dihydrobenzo[1,2-b:4,3-b']dipyrrole-6,2(3H)-diyl)carbonyl]]bis[8-(chloromethyl)-3,6,7,8-tetrahydro-1-methyl-benzo[1,2-b:4,3b']dipyrrol-4-ol(Compound 16).